Compounds that are S1P modulating agents and/or ATX modulating agents

ABSTRACT

Compounds of formula (I) can modulate the activity of one or more SIP receptors and/or the activity of autotaxin (ATX).

CLAIM OF PRIORITY RELATED APPLICATIONS

This application is a 35 U.S.C. § 371 national stage filing ofInternational Application No. PCT/US2013/053668, filed on Aug. 5, 2013,which claims priority to U.S. Provisional Application No. 61/679,984,filed on Aug. 6, 2012, the entire contents of which are incorporatedherein by reference in their entirety.

TECHNICAL FIELD

This invention relates to compounds that are S1P modulating agentsand/or ATX modulating agents, and methods of making and using suchcompounds.

BACKGROUND

Sphingosine 1-phosphate (S1P) is a lysophospholipid mediator that evokesa variety of cellular responses by stimulation of five members of theendothelial cell differentiation gene (EDG) receptor family. The EDGreceptors are G-protein coupled receptors (GPCRs) and on stimulationpropagate second messenger signals via activation of heterotrimericG-protein alpha (G_(α)) subunits and beta-gamma (G_(βγ)) dimers.Ultimately, this S1P-driven signaling results in cell survival,increased cell migration and, often, mitogenesis. The recent developmentof agonists targeting S1P receptors has provided insight regarding therole of this signaling system in physiologic homeostasis. For example,the immunomodulating agent, FTY720(2-amino-2-[2-(4-octylphenyl)ethyl]propane 1,3-diol), that followingphosphorylation, is an agonist at 4 of 5 S1P receptors, revealed thataffecting S1P receptor activity influences lymphocyte trafficking.Further, S1P type 1 receptor (S1P₁) antagonists cause leakage of thelung capillary endothelium, which suggests that S1P may be involved inmaintaining the integrity of the endothelial barrier in some tissuebeds. S1P type 4 receptors (S1P₄) are expressed mainly in leukocytes,and specifically S1P₄ mediates immunosuppressive effects of S1P byinhibiting proliferation and secretion of effector cytokines, whileenhancing secretion of the suppressive cytokine IL-10. See, for example,Wang, W. et. al., (2005) FASEB J. 19(12): 1731-3, which is incorporatedby reference in its entirety. S1P type 5 receptors (S1P₅) areexclusively expressed in oligodendrocytes and oligodendrocyte precursorcells (OPCs) and are vital for cell migration. Stimulation of S1P₅inhibits OPC migration, which normally migrate considerable distancesduring brain development. See, for example, Novgorodov, A. et al.,(2007) FASEB J, 21: 1503-1514, which is incorporated by reference in itsentirety.

S1P has been demonstrated to induce many cellular processes, includingthose that result in platelet aggregation, cell proliferation, cellmorphology, tumor-cell invasion, endothelial cell chemotaxis andangiogenesis. For these reasons, S1P receptors are good targets fortherapeutic applications such as wound healing, tumor growth inhibition,and autoimmune diseases.

Sphingosine-1-phosphate signals cells in part via a set of Gprotein-coupled receptors named S1P₁, S1P₂, S1P₃, S1P₄, and S1P₅(formerly EDG1, EDG5, EDG3, EDGE and EDGE). The EDG receptors areG-protein coupled receptors (GPCRs) and on stimulation propagate secondmessenger signals via activation of heterotrimeric G-protein alpha(G_(α)) subunits and beta-gamma (G_(βγ)) dimers. These receptors share50-55% amino acid sequence identity and cluster with three otherreceptors (LPA₁, LPA₂, and LPA₃ (formerly EDG2, EDG4 and EDG7) for thestructurally related lysophosphatidic acid (LPA).

A conformational shift is induced in the G-Protein Coupled Receptor(GPCR) when the ligand binds to that receptor, causing GDP to bereplaced by GTP on the α-subunit of the associated G-proteins andsubsequent release of the G-proteins into the cytoplasm. The α-subunitthen dissociates from the βγ-subunit and each subunit can then associatewith effector proteins, which activate second messengers leading to acellular response. Eventually the GTP on the G-proteins is hydrolyzed toGDP and the subunits of the G-proteins reassociate with each other andthen with the receptor. Amplification plays a major role in the generalGPCR pathway. The binding of one ligand to one receptor leads to theactivation of many G-proteins, each capable of associating with manyeffector proteins leading to an amplified cellular response.

S1P receptors make good drug targets because individual receptors areboth tissue and response specific. Tissue specificity of the S1Preceptors is desirable because development of an agonist or antagonistselective for one receptor localizes the cellular response to tissuescontaining that receptor, limiting unwanted side effects. Responsespecificity of the S1P receptors is also of importance because it allowsfor the development of agonists or antagonists that initiate or suppresscertain cellular responses without affecting other responses. Forexample, the response specificity of the S1P receptors could allow foran S1P mimetic that initiates platelet aggregation without affectingcell morphology.

Sphingosine-1-phosphate is formed as a metabolite of sphingosine in itsreaction with sphingosine kinase and is stored in abundance in theaggregates of platelets where high levels of sphingosine kinase existand sphingosine lyase is lacking. S1P is released during plateletaggregation, accumulates in serum, and is also found in malignantascites. Reversible biodegradation of S1P most likely proceeds viahydrolysis by ectophosphohydrolases, specifically thesphingosine-1-phosphate phosphohydrolases. Irreversible degradation ofS1P is catalyzed by S1P lyase yielding ethanolamine phosphate andhexadecenal.

Autotaxin (ATX, ENPP2) is a secreted glycoprotein widely present inbiological fluids, including blood, cancer ascites, synovial, pleuraland cerebrospinal fluids, originally isolated from the supernatant ofmelanoma cells as an autocrine motility stimulation factor (Stracke, M.L., et al. Identification, purification, and partial sequence analysisof autotaxin, a novel motility-stimulating protein. J Biol Chem 267,2524-2529 (1992), which is incorporated by reference in its entirety).ATX is encoded by a single gene on human chromosome 8 (mouse chromosome15) whose transcription, regulated by diverse transcription factors(Hoxal3, NFAT-1 and v-jun), results in four alternatively splicedisoforms (α, β, γ, and δ). See, for example, Giganti, A., et al Murineand Human Autotaxin alpha, beta, and gamma Isoforms: Gene organization,tissue distribution and biochemical characterization. J Biol Chem 283,7776-7789 (2008); and van Meeteren, L. A. & Moolenaar, W. H. Regulationand biological activities of the autotaxin-LPA axis. Prog Lipid Res 46,145-160 (2007); Hashimoto, et al, “Identification and BiochemicalCharaterization of a Novel Autotaxin Isoform, ATXδ,” J. of BiochemistryAdvance Access (Oct. 11, 2011); each of which is incorporated byreference in its entirety.

ATX is synthesized as a prepro-enzyme, secreted into the extracellularspace after the proteolytic removal of its N-terminal signal peptide(Jansen, S., el al Proteolytic maturation and activation of autotaxin(NPP2), a secreted metastasis-enhancing lysophospho lipase D. J Cell Sci118, 3081-3089 (2005), which is incorporated by reference in itsentirety). ATX is a member of the ectonucleotidepyrophosphatase/phosphodiesterase family of ectoenzymes (E-NPP) thathydrolyze phosphodiesterase (PDE) bonds of various nucleotides andderivatives (Stefan, C, Jansen, S. & Bollen, M. NPP-typeectophosphodiesterases: unity in diversity. Trends Biochem Sci 30,542-550 (2005), which is incorporated by reference in its entirety). Theenzymatic activity of ATX was enigmatic, until it was shown to beidentical to lysophospholipase D (lysoPLD) (Umezu-Goto, M., et al.Autotaxin has lysophospholipase D activity leading to tumor cell growthand motility by lysophosphatidic acid production. J Cell Biol 158,227-233 (2002), which is incorporated by reference in its entirety),which is widely present in biological fluids. Since ATX is aconstitutively active enzyme, the biological outcome of ATX action willlargely depend on its expression levels and the local availability ofits substrates. The major lysophospholipid substrate for ATX,lysophosphatidylcholine (LPC), is secreted by the liver and isabundantly present in plasma (at about 100 μM) as a predominantlyalbumin bound form (Croset, M., Brossard, N., Polette, A. & Lagarde, M.Characterization of plasma unsaturated lysophosphatidylcholines in humanand rat Biochem J 345 Pt 1, 61-67 (2000), which is incorporated byreference in its entirety). LPC is also detected in tumor-cellconditioned media (Umezu-Goto, M., et al.), presumably as a constituentof shed microvesicles. ATX, through its lysoPLD activity converts LPC tolysophosphatidic acid (LPA).

LPC is an important inflammatory mediator with recognized effects inmultiple cell types and pathophysiological processes. It is a majorcomponent of oxidized low density lipoprotein (oxLDL) and it can existin several other forms including free, micellar, bound to hydrophobicproteins such as albumin and incorporated in plasma membranes. It isproduced by the hydrolysis of phosphatidylcholine (PC) by PLA2 withconcurrent release of arachidonic acid and in turn of otherpro-inflammatory mediators (prostaglandins and leukotrienes). Moreover,LPC externalization constitutes a chemotactic signal to phagocyticcells, while interaction with its receptors can also stimulatelymphocytic responses. LPC has been shown to have therapeutic effects inexperimental sepsis, possibly by suppressing endotoxin-induced HMGB1release from macrophages/monocytes.

LPA, the product of ATX action on LPC, is a bioactive phospholipid withdiverse functions in almost every mammalian cell line (Moolenaar, W. H.,van Meeteren, L. A. & Giepmans, B. N. The ins and outs oflysophosphatidic acid signaling. Bioessays 28, 870-881 (2004), which isincorporated by reference in its entirety). LPA is a major constituentof serum bound tightly to albumin, gelsolin and possibly other as yetunidentified proteins. (See, e.g., Goetzl, E. J., et al Gelsolin bindingand. cellular presentation of lysophosphatidic acid. J Biol Chem 275,14573-14578 (2000); and Tigyi, G. & Miledi, R, Lysophosphatidates boundto serum albumin activate membrane currents in Xenopus oocytes andneurite retraction in PC12 pheochromocytoma cells. J Biol Chem 267,21360-21367 (1992); each of which is incorporated by reference in itsentirety.)

LPA is also found in other biofluids, such as saliva and follicularfluid, and has been implicated in a wide array of functions, such aswound heeling, tumor invasion and metastasis, neurogenesis, myelination,astrocytes outgrowth and neurite retraction. The long list of LPAfunctions was also explained with the discovery that it signals throughG-protein coupled receptors (GPCRs), via classical second messengerpathways. Five mammalian cell-surface LPA receptors have been identifiedso far. The best known are LPA1-3 (namely Edg-2, Edg-4 and Edg7) whichare all members of the so-called ‘endothelial differentiation gene’(EDG) family of GPCRs (Contos, J. J., Ishii, I. & Chun, J.Lysophosphatidic acid receptors. Mol Pharmacol 58, 1188-1196 (2000),which is incorporated by reference in its entirety). LPA receptors cancouple to at least three distinct G proteins (G_(q), G_(i) andG_(12/13)), which, in turn, feed into multiple effector systems. LPAactivates G_(q) and thereby stimulates phospholipase C (PLC), withsubsequent phosphatidylinositol-bisphosphate hydrolysis and generationof multiple second messengers leading to protein kinase C activation andchanges in cytosolic calcium. LPA also activates G_(i) which leads to atleast three distinct signaling routes: inhibition of adenylyl cyclasewith inhibition of cyclic AMP accumulation; stimulation of the mitogenicRAS-MAPK (mitogen-activated protein kinase) cascade; and activation ofphosphatidylinositol 3-kinase (PI3K), leading to activation of theguanosine diphosphate/guanosine triphosphate (GDP/GTP) exchange factorTIAM1 and the downstream RAC GTPase, as well as to activation of theAKT/PKB antiapoptotic pathway. Finally, LPA activates G_(12/13), leadingto activation of the small GTPase RhoA, which drives cytoskeletalcontraction and cell rounding. So, LPA not only signals via classicsecond messengers such as calcium, diacylglycerol and cAMP, but it alsoactivates RAS- and RHO-family GTPases, the master switches that controlcell proliferation, migration and morphogenesis.

LPA signaling through the RhoA-Rho kinase pathway mediates neuriteretraction and inhibition of axon growth. Interfering with LPA signalinghas been shown to promote axonal regeneration and functional recoveryafter CNS injury or cerebral ischemia. (See Broggini, et al., MolecularBiology of the Cell (2010), 21:521-537.) It has been reported thataddition of LPA to dorsal root fibers in ex vivo culture causesdemyelination, whereas LPC fails to cause significant demyelination ofnerve fibers in ex vivo cultures without further addition of recombinantATX to the culture which when added caused significant demyelination atequivalent levels to LPA presumable due to conversion of LPC to LPAthrough the enzymatic activity of ATX. Moreover, injury induceddemyelination was attenuated by about 50% in atx^(+/−) mice (Nagai, etal., Molecular Pain (2010), 6:78).

A number of diseases or disorders involve demyelination of the centralor peripheral nervous system which can occur for a number of reasonssuch as immune dysfunction as in multiple sclerosis, encephalomyelitis,Guillain-Barre Syndrome, chronic inflammatory demyelinatingpolyneuropathy (CIDP), transverse myelitis, and optic neuritis;demyelination due to injury such as spinal cord injury, traumatic braininjury, stroke, acute ischemic optic neuropathy, or other ischemia,cerebral palsy, neuropathy (e.g. neuropathy due to diabetes, chronicrenal failure, hypothyroidism, liver failure, or compression of thenerve (e.g. in Bell's palsy)), post radiation injury, and centralpontine myelolysis (CPM); inherited conditions such asCharcot-Marie-Tooth disease (CMT), Sjogren-Larsson syndrome, Refsumdisease, Krabbe disease, Canavan disease, Alexander disease,Friedreich's ataxia, Pelizaeus-Merzbacher disease, Bassen-Kornzweigsyndrome, metachromatic leukodystrophy (MLD), adrenoleukodystrophy, andnerve damage due to pernicious anemia; viral infection such asprogressive multifocal leukoencephalopathy (PML), Lyme disease, or tabesdorsalis due to untreated syphilis; toxic exposure due to chronicalcoholism (which is a possible cause of Marchiafava-Bignami disease),chemotherapy, or exposure to chemicals such as organophosphates; ordietary deficiencies such as vitamin B12 deficiency, vitamin Edeficiency and copper deficiency. Other demyelination disorders may haveunknown causes or multiple causes such as trigeminal neuralgia,Marchiafava-Bignami disease and Bell's palsy. One particularlysuccessful approach to treating demyelination disorders which are causedby autoimmune dysfunction has been to attempt to limit the extent ofdemyelination by treating the patient with immunoregulatory drugs.However, typically this approach has merely postponed but not avoidedthe onset of disability in these patients. Patients with demyelinationdue to other causes have even fewer treatment options. Therefore, theneed exists to develop new treatments for patients with demyelinationdiseases or disorders.

SUMMARY

A compound of formula (I), or a pharmaceutically acceptable saltthereof, can be an S1P modulating agent and/or an ATX modulating agent,e.g., an S1P4 antagonist or ATX inhibitor.

In one aspect, a compound is represented by formula (I):

or a pharmaceutically acceptable salt thereof.

X can be O, S(O)_(r), NR¹², C(O) or CH₂.

A¹, A², and A³ can each independently be CR² or N.

A⁶ and A⁷ can each independently be CR², C(R²)₂, N, or NR¹⁹.

One of A⁴ or A⁵ can be CR², C(R²)₂, N, or NR¹⁹, and the other can beCR⁴, provided that at least three of A¹, A², A³, A⁴, A⁵, A⁶ and A⁷ canindependently be CR² or C(R²)₂.

“------” can indicate a double or a single bond.

R¹ can be a C₆₋₂₀alkyl, a C₃₋₁₄carbocyclyl, a 3- to 15-memberedheterocyclyl, or a five- to 14-membered heteroaryl, wherein theheterocyclyl and the heteroaryl comprising from 1 to 10 heteroatomsindependently selected from N, S or O, and wherein R¹ may be optionallysubstituted with from one to six independently selected R⁶.

R², for each occurrence, can be independently selected from the groupconsisting of hydrogen, halo, hydroxyl, nitro, cyano, carboxy,C₁₋₆alkyl, C₁₋₆haloalkyl, C₃₋₈cycloalkyl, C₃₋₈halocycloalkyl,C₁₋₆alkoxy, C₁₋₆haloalkoxy, C₃₋₈cycloalkoxy, C₃₋₈halocycloalkoxy,C₁₋₆alkanoyl, amino, N—(C₁₋₆alkyl)amino, N,N-di-(C₁₋₆alkyl)amino,C₁₋₆alkoxycarbonyl, C₁₋₆alkanoyloxy, carbamoyl, N—(C₁₋₆alkyl)carbamoyl,N,N-di-(C₁₋₆alkyl)carbamoyl, C₁₋₆alkylamido, mercapto, C₁₋₆alkylthio,C₁₋₆alkylsulfonyl, sulfamoyl, N—(C₁₋₆alkyl)sulfamoyl,N,N-di-(C₁₋₆alkyl)sulfamoyl, and C₁₋₆alkylsulfonamido.

R⁴ can be a group represented by the following formula:

“

” can represent the point of attachment.

R⁵ can be a C₁₋₆alkylene, C₃₋₈carbocyclyl, a 3- to 8-memberedheterocyclyl, C₆₋₁₀aryl, a 5- to 10-membered heteroaryl, a bridged ringsystem comprising from 6 to 12 ring members, a spiro ring systemcomprising from 5-14 ring members; or a bicyclic ring system representedby the following formula:

where B′ and B″ can be independently selected from the group consistingof monocyclic C₃₋₈carbocyclyl, a monocyclic 3- to 8-memberedheterocyclyl, phenyl or a 5- to 6-membered heteroaryl; where R⁵ may beoptionally substituted with from 1 to 4 independently selected R¹¹.

R⁶, for each occurrence, can be independently selected from the groupconsisting of halo, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,C₃₋₈cycloalkyl, C₆₋₁₀aryl, C₁₋₆alkoxy-C₁₋₆alkyl, andtri-(C₁₋₆alkyl)silyl; or two R⁶ that are attached to the same carbonatom may form C₃₋₈spirocycloalkyl or 3- to 8-memberedspiroheterocycloalkyl.

R⁷ can be —C(O)OR¹⁵, —C(O)N(R¹⁶)₂, —C(O)N(R¹⁵)—S(O)₂R¹⁵, —S(O)₂OR¹⁵,—C(O)NHC(O)R¹⁵, —Si(O)OH, —B(OH)₂, —S(O)₂N(R¹⁵)₂, —O—P(O)(OR¹⁵)₂,—P(O)(OR¹⁵)₂, —CN, —S(O)₂NHC(O)R¹⁵, —C(O)NHS(O)₂R¹⁵, —C(O)NHOH,—C(O)NHCN, or a heteroaryl or a heterocyclyl selected from the groupconsisting of formulae (a)-(i′):

R⁸ and R⁹ can each independently be hydrogen, a carboxy, C₁₋₆alkyl, or aC₂₋₆alkenyl; or R⁸ and R⁹ together with the carbon to which they areattached can be —C(═O)—, a C₃₋₈spirocycloalkyl, or a 3- to 8-memberedspiroheterocycloalkyl.

R¹⁰ and R¹² can each, independently, be hydrogen or a C₁₋₆alkyl.

R¹¹, for each occurrence, can independently be halo, hydroxyl, nitro,cyano, C₁₋₆alkyl, C₁₋₆haloalkyl, C₁₋₆alkoxy, C₁₋₄haloalkoxy,C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₈cycloalkyl, C₃₋₈halocycloalkyl,C₃₋₈cycloalkoxy, C₃₋₈halocycloalkoxy, —NR^(a)R^(b), —C(O)NR^(a)R^(b),—N(R^(a))C(O)R^(b), —C(O)R^(a), —S(O)_(r)R^(a), or —N(R^(a))S(O)₂R^(b).

R¹⁵ for each occurrence can be independently selected from the groupconsisting of hydrogen, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₃₋₈cycloalkyl, C₃₋₈cycloalkenyl, C₆₋₁₀aryl, a 5 to 14 memberedheteroaryl, and a 3 to 15 membered heterocyclyl; where the heteroaryl orheterocyclyl can comprise from 1 to 10 heteroatoms independentlyselected from O, N, or S; and where R¹⁵ may be optionally substitutedwith from 1 to 3 substituents independently selected from the groupconsisting of halo, C₁₋₄alkoxy, C₁₋₄alkyl, cyano, nitro, hydroxyl,amino, N—(C₁₋₄alkyl)amino, N,N-di-(C₁₋₄alkyl)amino, carbamoyl,N—(C₁₋₄alkyl)carbamoyl, N,N-di-(C₁₋₄alkyl)carbamoyl, C₁₋₄alkylamido,C₁₋₄alkylsulfonyl, C₁₋₄alkylsulfonamido, sulfamoyl,N—(C₁₋₄alkyl)sulfamoyl, and N,N—(C₁₋₄dialkyl)-sulfamoyl.

R¹⁶ can be R¹⁵; or two R¹⁶ together with the nitrogen atom to which theyare attached form a 5 to 14 membered heteroaryl or a 3 to 15 memberedheterocyclyl, where the heteroaryl or heterocyclyl can comprise from 1to 10 heteroatoms independently selected from O, N, or S; and where theheteroaryl or heterocyclyl may be optionally substituted with from 1 to3 substituents independently selected from the group consisting of halo,C₁₋₄alkoxy, C₁₋₄alkyl, cyano, nitro, hydroxyl, amino,N—(C₁₋₄alkyl)amino, N,N-di-(C₁₋₄alkyl)amino, carbamoyl,N—(C₁₋₄alkyl)carbamoyl, N,N-di-(C₁₋₄alkyl)carbamoyl, C₁₋₄alkylamido,C₁₋₄alkylsulfonyl, C₁₋₄alkylsulfonamido, sulfamoyl,N—C₁₋₄alkylsulfamoyl, and N,N—(C₁₋₄dialkyl)-sulfamoyl.

R¹⁷ and R¹⁸, for each occurrence, can each independently be hydrogen, ahalo, or a C₁₋₄haloalkyl.

R¹⁹ for each occurrence can be independently selected from the groupconsisting of hydrogen, carboxy, C₁₋₆alkyl, C₁₋₆haloalkyl,C₃₋₈cycloalkyl, C₃₋₈halocycloalkyl, C₁₋₆alkanoyl, C₁₋₆alkoxycarbonyl,carbamoyl, N—(C₁₋₆alkyl)carbamoyl, N,N-di-(C₁₋₆alkyl)carbamoyl,C₁₋₆alkylsulfonyl, sulfamoyl, N—(C₁₋₆alkyl)sulfamoyl, andN,N-di-(C₁₋₆alkyl)sulfamoyl.

R^(a) and R^(b), for each occurrence, can independently be hydrogen,C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₈cycloalkyl, C₆₋₁₀aryl, orC₃₋₈halocycloalkyl.

R^(c) is hydrogen of a C₁₋₄alkyl.

m can be 0 or 1, provided that when m is 0, R⁵ can comprise at least onenitrogen.

p can be 0 or an integer from 1 to 6.

r, for each occurrence, can independently be 0, 1, or 2.

In some embodiments, R⁵ can be substituted by —(CR¹⁷R¹⁸)_(p)—R⁷ and canbe optionally substituted by from 1 to 3 independently selected R¹¹.

In some embodiments, the compound can be represented by structuralformula (II):

In some embodiments, the compound can be represented by structuralformula (III):

In some embodiments, R¹ can be a cyclohexyl which is optionallysubstituted with from one to three independently selected R⁶.

In some embodiments, m can be 0; and R⁵ can be selected from the groupconsisting of:

In some embodiments, m can be 1; and R⁵ can be cyclobutyl, cyclopentylor cyclohexyl each of which may be optionally substituted with from 1 to3 independently selected R¹¹.

In some embodiments, R⁷ can be COOH.

In some embodiments, A¹, A², and A³ can each independently be CR².

In some embodiments, A⁴ can be CR⁴, A⁵ can be CR², and A⁷ can be CR²; orA⁵ can be CR⁴, and A⁷ can be CR².

In some embodiments, A¹, A², A³, A⁶, and A⁷ can each independently beCR², and one of A⁴ or A⁵ can be CR² and the other can be CR⁴.

In some embodiments, X can be O.

In another aspect, a compound can be selected from the group consistingof:

-   8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylic    acid;-   9-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylic    acid;-   3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylic    acid;-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   2-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)octahydrocyclopenta[c]pyrrole-5-carboxylic    acid;-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)pyrrolidine-3-carboxylic    acid;-   3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)cyclobutanecarboxylic    acid;-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azepane-4-carboxylic    acid;-   1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((cis-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((cis-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((cis-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((4,4-dimethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-phenylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-butylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-cyclopentylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-(heptyloxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   9-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylic    acid;-   8-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylic    acid;-   1-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-(heptyloxy)naphthalen-1-yl)methyl)piperidine-4-carboxylic    acid; and-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylic    acid;

or a pharmaceutically acceptable salt thereof.

In another aspect, a pharmaceutical composition includes apharmaceutically acceptable carrier or excipient and a compound offormula (I), or a pharmaceutically acceptable salt thereof.

In another aspect, a method of preventing, treating, or reducingsymptoms of a condition mediated by S1P activity or ATX activity in amammal includes administering to said mammal an effective amount of acompound of formula (I), or a pharmaceutically acceptable salt thereof.

The condition can be selected from the group consisting of multiplesclerosis, an autoimmune disease, a chronic inflammatory disorder,asthma, an inflammatory neuropathy, arthritis, transplantationrejection, Crohn's disease, ulcerative colitis, lupus erythematosis,psoriasis, an ischemia-reperfusion injury, a solid tumor, a tumormetastasis, a disease associated with angiogenesis, a vascular disease,a pain condition, an acute viral disease, an inflammatory bowelcondition, insulin-dependent diabetes, non-insulin dependent diabetes, afibrosis of the lung, or a malignancy of the lung in a mammal.

In some embodiments, the condition can be multiple sclerosis.

In some embodiments, the condition can be rheumatoid arthritis.

The method can further include administering to said mammal an effectiveamount of one or more drugs selected from the group consisting of: acorticosteroid, a bronchodilator, an antiasthmatic, an antiinflammatory,an antirheumatic, an immunosuppressant, an antimetabolite, animmunomodulating agent, an antipsoriatic, and an antidiabetic.

In another aspect, a method of preventing, treating, or reducing chronicpain in a mammal includes administering to said mammal an effectiveamount of a compound of formula (I), or a pharmaceutically acceptablesalt thereof.

The chronic pain can be inflammatory pain.

The chronic pain can be neuropathic pain.

Other features or advantages will be apparent from the followingdetailed description of several embodiments, and also from the appendedclaims.

DETAILED DESCRIPTION

The disclosed compounds can be S1P modulating agents and/or ATXmodulating agents. In other words, the disclosed compounds can haveactivity as receptor agonists or receptor antagonists at one or more S1Preceptors, or as an ATX modulating agent. In particular, the compoundscan be S1P4 antagonists, or ATX inhibitors. A given compound can be anS1P modulating agent with little or substantially no ATX activity; orcan be an ATX modulating agent with little or substantially no S1Pactivity; or, in some cases, can simultaneously be an S1P modulatingagent and an ATX modulating agent. Preferably, a given compound iseither an S1P modulating agent with little or substantially no ATXactivity; or is an ATX modulating agent with little or substantially noS1P activity.

A compound, or a pharmaceutically acceptable salt thereof, can berepresented by formula (I):

or a pharmaceutically acceptable salt thereof.

X can be 0, S(O)_(r), NR¹², C(O) or CH₂.

A¹, A², and A³ can each independently be CR² or N.

A⁶ and A⁷ can each independently be CR², C(R²)₂, N, or NR¹⁹.

One of A⁴ or A⁵ can be CR², C(R²)₂, N, or NR¹⁹, and the other can beCR⁴, provided that at least three of A¹, A², A³, A⁴, A⁵, A⁶ and A⁷ canindependently be CR² or C(R²)₂.

“------” can indicate a double or a single bond.

R¹ can be a C₆₋₂₀alkyl, a C₃₋₁₄carbocyclyl, a 3- to 15-memberedheterocyclyl, or a five- to 14-membered heteroaryl, wherein theheterocyclyl and the heteroaryl comprising from 1 to 10 heteroatomsindependently selected from N, S or O, and wherein R¹ may be optionallysubstituted with from one to six independently selected R⁶.

R², for each occurrence, can be independently selected from the groupconsisting of hydrogen, halo, hydroxyl, nitro, cyano, carboxy,C₁₋₆alkyl, C₁₋₆haloalkyl, C₃₋₈cycloalkyl, C₃₋₈halocycloalkyl,C₁₋₆alkoxy, C₁₋₆haloalkoxy, C₃₋₈cycloalkoxy, C₃₋₈halocycloalkoxy,C₁₋₆alkanoyl, amino, N—(C₁₋₆alkyl)amino, N,N-di-(C₁₋₆alkyl)amino,C₁₋₆alkoxycarbonyl, C₁₋₆alkanoyloxy, carbamoyl, N—(C₁₋₆alkyl)carbamoyl,N,N-di-(C₁₋₆alkyl)carbamoyl, C₁₋₆alkylamido, mercapto, C₁₋₆alkylthio,C₁₋₆alkylsulfonyl, sulfamoyl, N—(C₁₋₆alkyl)sulfamoyl,N,N-di-(C₁₋₆alkyl)sulfamoyl, and C₁₋₆alkylsulfonamido.

R⁴ can be a group represented by the following formula:

“

” can represent the point of attachment.

R⁵ can be a C₁₋₆alkylene, C₃₋₈carbocyclyl, a 3- to 8-memberedheterocyclyl, C₆₋₁₀aryl, a 5- to 10-membered heteroaryl, a bridged ringsystem comprising from 6 to 12 ring members, a spiro ring systemcomprising from 5-14 ring members; or a bicyclic ring system representedby the following formula:

where B′ and B″ can be independently selected from the group consistingof monocyclic C₃₋₈carbocyclyl, a monocyclic 3- to 8-memberedheterocyclyl, phenyl or a 5- to 6-membered heteroaryl; where R⁵ may beoptionally substituted with from 1 to 4 independently selected R¹¹.

R⁶, for each occurrence, can be independently selected from the groupconsisting of halo, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,C₃₋₈cycloalkyl, C₆₋₁₀aryl, C₁₋₆alkoxy-C₁₋₆alkyl, andtri-(C₁₋₆alkyl)silyl; or two R⁶ that are attached to the same carbonatom may form C₃₋₈spirocycloalkyl or 3- to 8-memberedspiroheterocycloalkyl.

R⁷ can be —C(O)OR¹⁵, —C(O)N(R¹⁶)₂, —C(O)N(R¹⁵)—S(O)₂R¹⁵, —S(O)₂OR¹⁵,—C(O)NHC(O)R¹⁵, —Si(O)OH, —B(OH)₂, —S(O)₂N(R¹⁵)₂, —O—P(O)(OR¹⁵)₂,—P(O)(OR¹⁵)₂, —CN, —S(O)₂NHC(O)R¹⁵, —C(O)NHS(O)₂R¹⁵, —C(O)NHOH,—C(O)NHCN, or a heteroaryl or a heterocyclyl selected from the groupconsisting of formulae (a)-(i′):

R⁸ and R⁹ can each independently be hydrogen, a carboxy, C₁₋₆alkyl, or aC₂₋₆alkenyl; or R⁸ and R⁹ together with the carbon to which they areattached can be —C(═O)—, a C₃₋₈spirocycloalkyl, or a 3- to 8-memberedspiroheterocycloalkyl.

R¹⁰ and R¹² can each, independently, be hydrogen or a C₁₋₆alkyl.

R¹¹, for each occurrence, can independently be halo, hydroxyl, nitro,cyano, C₁₋₆alkyl, C₁₋₆haloalkyl, C₁₋₆alkoxy, C₁₋₄haloalkoxy,C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₈cycloalkyl, C₃₋₈halocycloalkyl,C₃₋₈cycloalkoxy, C₃₋₈halocycloalkoxy, —NR^(a)R^(b), —C(O)NR^(a)R^(b),—N(R^(a))C(O)R^(b), —C(O)R^(a), —S(O)_(r)R^(a), or —N(R^(a))S(O)₂R^(b).

R¹⁵ for each occurrence can be independently selected from the groupconsisting of hydrogen, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₃₋₈cycloalkyl, C₃₋₈cycloalkenyl, C₆₋₁₀aryl, a 5 to 14 memberedheteroaryl, and a 3 to 15 membered heterocyclyl; where the heteroaryl orheterocyclyl can comprise from 1 to 10 heteroatoms independentlyselected from O, N, or S; and where R¹⁵ may be optionally substitutedwith from 1 to 3 substituents independently selected from the groupconsisting of halo, C₁₋₄alkoxy, C₁₋₄alkyl, cyano, nitro, hydroxyl,amino, N—(C₁₋₄alkyl)amino, N,N-di-(C₁₋₄alkyl)amino, carbamoyl,N—(C₁₋₄alkyl)carbamoyl, N,N-di-(C₁₋₄alkyl)carbamoyl, C₁₋₄alkylamido,C₁₋₄alkylsulfonyl, C₁₋₄alkylsulfonamido, sulfamoyl,N—(C₁₋₄alkyl)sulfamoyl, and N,N—(C₁₋₄dialkyl)-sulfamoyl.

R¹⁶ can be R¹⁵; or two R¹⁶ together with the nitrogen atom to which theyare attached form a 5 to 14 membered heteroaryl or a 3 to 15 memberedheterocyclyl, where the heteroaryl or heterocyclyl can comprise from 1to 10 heteroatoms independently selected from O, N, or S; and where theheteroaryl or heterocyclyl may be optionally substituted with from 1 to3 substituents independently selected from the group consisting of halo,C₁₋₄alkoxy, C₁₋₄alkyl, cyano, nitro, hydroxyl, amino,N—(C₁₋₄alkyl)amino, N,N-di-(C₁₋₄alkyl)amino, carbamoyl,N—(C₁₋₄alkyl)carbamoyl, N,N-di-(C₁₋₄alkyl)carbamoyl, C₁₋₄alkylamido,C₁₋₄alkylsulfonyl, C₁₋₄alkylsulfonamido, sulfamoyl,N—C₁₋₄alkylsulfamoyl, and N,N—(C₁₋₄dialkyl)-sulfamoyl.

R¹⁷ and R¹⁸, for each occurrence, can each independently be hydrogen, ahalo, or a C₁₋₄haloalkyl.

R¹⁹ for each occurrence can be independently selected from the groupconsisting of hydrogen, carboxy, C₁₋₆alkyl, C₁₋₆haloalkyl,C₃₋₈cycloalkyl, C₃₋₈halocycloalkyl, C₁₋₆alkanoyl, C₁₋₆alkoxycarbonyl,carbamoyl, N—(C₁₋₆alkyl)carbamoyl, N,N-di-(C₁₋₆alkyl)carbamoyl,C₁₋₆alkylsulfonyl, sulfamoyl, N—(C₁₋₆alkyl)sulfamoyl, andN,N-di-(C₁₋₆alkyl)sulfamoyl.

R^(a) and R^(b), for each occurrence, can independently be hydrogen,C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₈cycloalkyl, C₆₋₁₀aryl, orC₃₋₈halocycloalkyl.

R^(c) is hydrogen or a C₁₋₄alkyl.

m can be 0 or 1, provided that when m is 0, R⁵ can comprise at least onenitrogen.

p can be 0 or an integer from 1 to 6.

r, for each occurrence, can independently be 0, 1, or 2.

In some embodiments, R⁵ can be substituted by —(CR¹⁷R¹⁸)_(p)—R⁷ and canbe optionally substituted by from 1 to 3 independently selected R¹¹.

In some embodiments, the compound can be represented by structuralformula (II):

In some embodiments, the compound can be represented by structuralformula (III):

In some embodiments, R¹ can be a cyclohexyl which is optionallysubstituted with from one to three independently selected R⁶.

In some embodiments, m can be 0; and R⁵ can be selected from the groupconsisting of:

In some embodiments, m can be 1; and R⁵ can be cyclobutyl, cyclopentylor cyclohexyl each of which may be optionally substituted with from 1 to3 independently selected R¹¹.

In some embodiments, R⁷ can be COOH.

In some embodiments, A¹, A², and A³ can each independently be CR².

In some embodiments, A⁴ can be CR⁴, A⁵ can be CR², and A⁷ can be CR²; orA⁵ can be CR⁴, and A⁷ can be CR².

In some embodiments, A¹, A², A³, A⁶, and A⁷ can each independently beCR², and one of A⁴ or A⁵ can be CR² and the other can be CR⁴.

In some embodiments, X can be O.

A compound can be selected from the group consisting of:

-   8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylic    acid;-   9-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylic    acid;-   3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylic    acid;-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   2-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)octahydrocyclopenta[c]pyrrole-5-carboxylic    acid;-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)pyrrolidine-3-carboxylic    acid;-   3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)cyclobutanecarboxylic    acid;-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azepane-4-carboxylic    acid;-   1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((cis-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((cis-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((cis-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((4,4-dimethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-phenylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-butylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-cyclopentylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylic    acid;-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-(heptyloxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   9-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylic    acid;-   8-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylic    acid;-   1-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic    acid;-   1-((5-(heptyloxy)naphthalen-1-yl)methyl)piperidine-4-carboxylic    acid; and-   1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylic    acid;

or a pharmaceutically acceptable salt thereof.

The term “fused ring system,” as used herein, is a ring system that hastwo or three rings (preferably two rings) independently selected fromcarbocyclyl, heterocyclyl, aryl or heteroaryl rings that share one side.A fused ring system may have from 4-15 ring members, preferably from5-10 ring members. Examples of fused ring systems includeoctahydroisoquinolin-2(1H)-yl, 2,3-dihydro-1H-indenyl,octahydro-1H-pyrido[1,2-a]pyrazinyl, and decahydroisoquinolinyl).

The term “bridged ring system,” as used herein, is a ring system thathas a carbocyclyl or heterocyclyl ring wherein two non-adjacent atoms ofthe ring are connected (bridged) by one or more (preferably from one tothree) atoms selected from C, N, O, or S. A bridged ring system can havemore than one bridge within the ring system (e.g., adamantyl). A bridgedring system may have from 6-10 ring members, preferably from 7-10 ringmembers. Examples of bridged ring systems include adamantyl,9-azabicyclo[3.3.1]nonan-9-yl, 8-azabicyclo[3.2.1]octanyl,bicyclo[2.2.2]octanyl, 3-azabicyclo[3.1.1]heptanyl,bicyclo[2.2.1]heptanyl, (1R,5S)-bicyclo[3.2.1]octanyl,3-azabicyclo[3.3.1]nonanyl, and bicyclo[2.2.1]heptanyl. More preferably,the bridged ring system is selected from the group consisting of9-azabicyclo[3.3.1]nonan-9-yl, 8-azabicyclo[3.2.1]octanyl, andbicyclo[2.2.2]octanyl.

The term “spiro ring system,” as used herein, is a ring system that hastwo rings each of which are independently selected from a carbocyclyl ora heterocyclyl, wherein the two ring structures having one atom incommon. Spiro ring systems have from 5 to 14 ring members. Example ofspiro ring systems include 2-azaspiro[3.3]heptanyl, spiropentanyl,2-oxa-6-azaspiro[3.3]heptanyl, 2,7-diazaspiro[3.5]nonanyl,2-oxa-7-azaspiro[3.5]nonanyl, 6-oxa-9-azaspiro[4.5]decanyl,6-oxa-2-azaspiro[3.4]octanyl, 5-azaspiro[2.3]hexanyl and2,8-diazaspiro[4.5]decanyl.

As used herein, the term “alkyl” refers to a fully saturated branched orunbranched hydrocarbon moiety. Preferably the alkyl comprises 1 to 20carbon atoms, more preferably 1 to 16 carbon atoms, 1 to 10 carbonatoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. In some embodiments,an alkyl comprises from 6 to 20 carbon atoms. Representative examples ofalkyl include, but are not limited to, methyl, ethyl, n-propyl,iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl,isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl,2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, or n-decyl.

“Alkylene” refers to a divalent alkyl group. Examples of alkylene groupsinclude methylene, ethylene, propylene, n-butylene, and the like. Thealkylene is attached to the rest of the molecule through a single bondand to the radical group through a single bond. The points of attachmentof the alkylene to the rest of the molecule and to the radical group canbe through one carbon or any two carbons within the carbon chain.

As used herein, the term “haloalkyl” refers to an alkyl, as definedherein, that is substituted by one or more halo groups as definedherein. Preferably the haloalkyl can be monohaloalkyl, dihaloalkyl orpolyhaloalkyl including perhaloalkyl. A monohaloalkyl can have one iodo,bromo, chloro or fluoro substituent. Dihaloalkyl and polyhaloalkylgroups can be substituted with two or more of the same halo atoms or acombination of different halo groups. Non-limiting examples of haloalkylinclude fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl,dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl,difluorochloromethyl, dichlorofluoromethyl, difluoroethyl,difluoropropyl, dichloroethyl and dichloropropyl. A perhaloalkyl refersto an alkyl having all hydrogen atoms replaced with halo atoms.Preferred haloalkyl groups are trifluoromethyl and difluoromethyl.

“Halogen” or “halo” may be fluoro, chloro, bromo or iodo.

As used herein, the term “alkoxy” refers to alkyl-O—, wherein alkyl isdefined herein above. Representative examples of alkoxy include, but arenot limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy,tert-butoxy, pentyloxy, hexyloxy, cyclopropyloxy-, cyclohexyloxy- andthe like. Preferably, alkoxy groups have about 1-6 carbon atoms, morepreferably about 1-4 carbon atoms.

As used herein, the term “haloalkoxy” refers to haloalkyl-O—, whereinhaloalkyl is defined herein above. Representative example of haloalkoxygroups are trifluoromethoxy, difluoromethoxy, and 1,2-dichloroethoxy.Preferably, haloalkoxy groups have about 1-6 carbon atoms, morepreferably about 1-4 carbon atoms.

As used herein, the term “alkylthio” refers to alkyl-S—, wherein alkylis defined herein above.

As used herein, the term “carbocyclyl” refers to saturated or partiallyunsaturated (but not aromatic) monocyclic, bicyclic or tricyclichydrocarbon groups of 3-14 carbon atoms, preferably 3-9, or morepreferably 3-8 carbon atoms. Carbocyclyls include fused or bridged ringsystems. The term “carbocyclyl” encompasses cycloalkyl groups. The term“cycloalkyl” refers to completely saturated monocyclic, bicyclic ortricyclic hydrocarbon groups of 3-12 carbon atoms, preferably 3-9, ormore preferably 3-8 carbon atoms. Exemplary monocyclic carbocyclylgroups include, but are not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclopentenyl, cyclohexyl or cyclohexenyl. Exemplarybicyclic carbocyclyl groups include bornyl, decahydronaphthyl,bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl,6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl,or bicyclo[2.2.2]octyl. Exemplary tricyclic carbocyclyl groups includeadamantyl.

As used herein, the term “halocycloalkyl” refers to a cycloalkyl, asdefined herein, that is substituted by one or more halo groups asdefined herein. Preferably the halocycloalkyl can be monohalocycloalkyl,dihalocycloalkyl or polyhalocycloalkyl including perhalocycloalkyl. Amonohalocycloalkyl can have one iodo, bromo, chloro or fluorosubstituent. Dihalocycloalkyl and polyhalocycloalkyl groups can besubstituted with two or more of the same halo atoms or a combination ofdifferent halo groups.

As used herein, the term “cycloalkoxy” refers to cycloalkyl-O—, whereincycloalkyl is defined herein above.

As used herein, the term “halocycloalkoxy” refers to halocycloalkyl-O—,wherein halocycloalkyl is defined herein above.

The term “spirocycloalkyl” as used herein, is a cycloalkyl that has onering atom in common with the group to which it is attached.Spirocycloalkyl groups may have from 3 to 14 ring members. In apreferred embodiment, the spirocycloalkyl has from 3 to 8 ring carbonatoms and is monocyclic.

The term “aryl” refers to monocyclic, bicyclic or tricyclic aromatichydrocarbon groups having from 6 to 14 carbon atoms in the ring portion.In one embodiment, the term aryl refers to monocyclic and bicyclicaromatic hydrocarbon groups having from 6 to 10 carbon atoms.Representative examples of aryl groups include phenyl, naphthyl,fluorenyl, and anthracenyl.

The term “aryl” also refers to a bicyclic or tricyclic group in which atleast one ring is aromatic and is fused to one or two non-aromatichydrocarbon ring(s). Nonlimiting examples include tetrahydronaphthalene,dihydronaphthalenyl and indanyl.

As used herein, the term “heterocyclyl” refers to a saturated orunsaturated, non-aromatic monocyclic, bicyclic or tricyclic ring systemwhich has from 3- to 15-ring members at least one of which is aheteroatom, and up to 10 of which may be heteroatoms, wherein theheteroatoms are independently selected from O, S and N, and wherein Nand S can be optionally oxidized to various oxidation states. In oneembodiment, a heterocyclyl is a 3-8-membered monocyclic. In anotherembodiment, a heterocyclyl is a 6-12-membered bicyclic. In yet anotherembodiment, a heterocyclycyl is a 10-15-membered tricyclic ring system.The heterocyclyl group can be attached at a heteroatom or a carbon atom.Heterocyclyls include fused or bridged ring systems. The term“heterocyclyl” encompasses heterocycloalkyl groups. The term“heterocycloalkyl” refers to completely saturated monocyclic, bicyclicor tricyclic heterocyclyl comprising 3-15 ring members, at least one ofwhich is a heteroatom, and up to 10 of which may be heteroatoms, whereinthe heteroatoms are independently selected from O, S and N, and whereinN and S can be optionally oxidized to various oxidation states. Examplesof heterocyclyls include dihydrofuranyl, [1,3]dioxolane, 1,4-dioxane,1,4-dithiane, piperazinyl, 1,3-dioxolane, imidazolidinyl, imidazolinyl,pyrrolidine, dihydropyran, oxathiolane, dithiolane, 1,3-dioxane,1,3-dithianyl, oxathianyl, thiomorpholinyl, oxiranyl, aziridinyl,oxetanyl, azetidinyl, tetrahydrofuranyl, pyrrolidinyl,tetrahydropyranyl, piperidinyl, morpholinyl, piperazinyl, azepinyl,oxapinyl, oxazepinyl and diazepinyl.

The term “spiroheterocycloalkyl” as used herein, is a heterocycloalkylthat has one ring atom in common with the group to which it is attached.Spiroheterocycloalkyl groups may have from 3 to 15 ring members. In apreferred embodiment, the spiroheterocycloalkyl has from 3 to 8 ringatoms selected from carbon, nitrogen, sulfur and oxygen and ismonocyclic.

As used herein, the term “heteroaryl” refers to a 5-14 memberedmonocyclic-, bicyclic-, or tricyclic-ring system, having 1 to 10heteroatoms independently selected from N, O or S, wherein N and S canbe optionally oxidized to various oxidation states, and wherein at leastone ring in the ring system is aromatic. In one embodiment, theheteroaryl is monocyclic and has 5 or 6 ring members. Examples ofmonocyclic heteroaryl groups include pyridyl, thienyl, furanyl,pyrrolyl, pyrazolyl, imidazoyl, oxazolyl, isoxazolyl, thiazolyl,isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl and tetrazolyl. Inanother embodiment, the heteroaryl is bicyclic and has from 8 to 10 ringmembers. Examples of bicyclic heteroaryl groups include indolyl,benzofuranyl, quinolyl, isoquinolyl indazolyl, indolinyl, isoindolyl,indolizinyl, benzamidazolyl, quinolinyl, 5,6,7,8-tetrahydroquinoline and6,7-dihydro-5H-pyrrolo[3,2-d]pyrimidine.

An amino is a group having the formula NH₂—. The term N-alkylamino is anamino group in which one of the hydrogen atoms is replaced with an alkylgroup. The term N,N-dialkylamino is an amino group in which eachhydrogen atoms is replaced with an alkyl group which may be the same ordifferent.

The term “alkanoyl” refers to alkyl-C(O)— wherein the alkyl is definedas above.

The term “alkoxycarbonyl” refers to alkoxy-C(O)—, wherein the alkoxygroup is defined as above.

The term “alkanoyloxy” refers to alkyl-C(O)O—, wherein the alkyl isdefined as above.

A carbamoyl is a group having the formula NH₂C(O)—. The termN-alkylcarbamoyl is a carbamoyl group in which one of the hydrogen atomsis replaced with an alkyl group. The term N,N-dialkylcarbamoyl is acarbamoyl group in which each hydrogen atoms is replaced with an alkylgroup which may be the same or different.

The term “alkylamido” refers to a group having the formulaalkyl-C(O)—NH—. As used herein, the term “alkylsulfonyl” refers to agroup having the formula alkyl-SO₂—.

A sulfamoyl is a group having the formula NH₂S(O)₂—. The termN-alkylsulfamoyl is a sulfamoyl group in which one of the hydrogen atomsis replaced with an alkyl group. The term N,N-dialkylsulfamoyl is asulfamoyl group in which each hydrogen atoms is replaced with an alkylgroup which may be the same or different.

The term “alkylsulfonamido” refers to a group having the formulaalkyl-S(O)₂—NH—.

The term “trialkylsilyl” refers to (alkyl)₃-Si—, wherein each of thealkyl groups may be the same or different.

The number of carbon atoms in a group is specified herein by the prefix“C_(x-xx)”, wherein x and xx are integers. For example, “C₁₋₄alkyl” isan alkyl group which has from 1 to 4 carbon atoms; C₁₋₆alkoxy is analkoxy group having from 1 to 6 carbon atoms; C₆₋₁₀aryl is an aryl groupwhich has from 6 to 10 carbon atoms; C₁₋₄haloalkyl is a haloalkyl groupwhich has from 1 to 4 carbon atoms; and N,N-di-C₁₋₆alkylamino is aN,N-dialkylamino group in which the nitrogen is substituted with twoalkyl groups each of which is independently from 1 to 6 carbon atoms.

The phrase “compound of the invention,” as used herein, refers tocompounds represented by formulae (I), (II), and (III), and any of thespecific examples disclosed herein.

The disclosed compounds can contain one or more asymmetric centers inthe molecule. In accordance with the present disclosure any structurethat does not designate the stereochemistry is to be understood asembracing all the various optical isomers (e.g., diastereomers andenantiomers) in pure or substantially pure form, as well as mixturesthereof (such as a racemic mixture, or an enantiomerically enrichedmixture). It is well known in the art how to prepare such opticallyactive forms (for example, resolution of the racemic form byrecrystallization techniques, synthesis from optically-active startingmaterials, by chiral synthesis, or chromatographic separation using achiral stationary phase). The compounds can be isotopically-labeledcompounds, for example, compounds including various isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, iodine, orchlorine. The disclosed compounds may exist in tautomeric forms andmixtures and separate individual tautomers are contemplated. Inaddition, some compounds may exhibit polymorphism.

By way of clarity, compounds of the invention included all isotopes ofthe atoms present in formulae (I), (II), and (III), and any of theexamples or embodiments disclosed herein. For example, H (or hydrogen)represents any isotopic form of hydrogen including ¹H, ²H (D), and ³H(T); C represents any isotopic form of carbon including and ¹²C, ¹³C,and ¹⁴C; O represents any isotopic form of oxygen including ¹⁶O, ¹⁷O and¹⁸O; N represents any isotopic form of nitrogen including ¹³N, ¹⁴N and¹⁵N; P represents any isotopic form of phosphorous including ³¹P and³²P; S represents any isotopic form of sulfur including ³²S and ³⁵S; Frepresents any isotopic form of fluorine including ¹⁹F and ¹⁸F; Clrepresents any isotopic form of chlorine including ³⁵Cl, ³⁷Cl and ³⁶Cl;and the like. In a preferred embodiment, compounds represented byformulae (I), (II), and (III), and any of the examples or embodimentsdisclosed herein comprises isotopes of the atoms therein in theirnaturally occurring abundance. However, in certain instances, it isdesirable to enrich one or more atom in a particular isotope which wouldnormally be present in less abundance. For example, ¹H would normally bepresent in greater than 99.98% abundance; however, a compound of theinvention can be enriched in ²H or ³H at one or more positions where His present. In particular embodiments of the compounds of formula (I),when, for example, hydrogen is enriched in the deuterium isotope, thesymbol “D” may be used to represent the enrichment in deuterium. In oneembodiment, when a compound of the invention is enriched in aradioactive isotope, for example ³H and ¹⁴C, they may be useful in drugand/or substrate tissue distribution assays. It is to be understood thatthe invention encompasses all such isotopic forms which modulate ATXactivity.

Compounds of the invention can modulate the activity of S1P receptors. Acompound of the invention can have S1P receptor agonist or antagonistactivity. The compound can be selective for the S1P4 receptor. Thecompound can be a selective S1P4 antagonist. Being selective can meanthat the compound binds to the receptor (or relatively small group ofrelated molecules or proteins) in a complex mixture, or in other words,when exposed to a variety of closely related receptor types, thecompound can bind preferentially to just one of the receptor types.

The compound can have a greater affinity for the S1P4 receptor, by at byat least 100-fold, by at least 50-fold, by at least 10-fold, by at least5-fold or by at least 2-fold, than for S1P1 receptor, S1P2 receptor,S1P3 receptor, or S1P5 receptor.

An inhibitor of S1P4 mediated activity can block S1P interaction with anS1P4 receptor. For example, the inhibitor can be an antagonist of anS1P4 receptor. An antagonist can be a molecule that has affinity for thereceptor but does not induce activity or a specific activity from thereceptor. The antagonist can bind with an S1P4 receptor with an IC₅₀value of less than 1 μM, less than 750 nM, less than 500 nM, less than250 nM or less than 100 nM. The antagonist can bind with an S1P4receptor with an IC₅₀ value in a range between 1 nM and 1 μM, between 1nM and 500 nM, between 10 nM and 250 nM, between 25 nm and 100 nM, orbetween 50 nM and 100 nM.

The compounds can also promote oligodendrocyte progenitor celldifferentiation. The compounds can promote myelination or remyelination.

An “S1P modulating agent” refers a compound or composition that iscapable of inducing a detectable change in S1P receptor activity in vivoor in vitro (e.g., at least 10% increase or decrease in S1P activity asmeasured by a given assay such as the assays described in the examplesand known in the art. “S1P receptor,” refers to all of the S1P receptorsubtypes (for example, the S1P receptors S1P1, S1P2, S1P3, S1P4, orS1P5), unless the specific subtype is indicated. It is well known in theart how to determine S1P agonist or antagonist activity using thestandard tests described herein, or using other similar tests which arewell known in the art. In some cases, depending on the cell type andconditions used, an S1P modulating agent can have agonist or antagonistactivity, even at the same receptor subtype.

The biological effects of an S1P modulating agent vary depending onwhether the compound has S1P receptor agonist or antagonist activity.Potential uses of an S1P modulating agent include, but are not limitedto, prevention or treatment of a pathological condition or symptom in amammal. For example, the condition can include asthma, an inflammatoryneuropathies, arthritis, lupus erythematosis, psoriasis, an ischemiareperfusion injury, a solid tumor, a tumor metastasis, a diseaseassociated with angiogenesis, a vascular disease, a pain condition, anacute viral disease, or insulin-dependent diabetes, and non-insulindependent diabetes. The condition can alter lymphocyte trafficking as amethod of treatment for neuropathic pain, inflammation-induced pain(e.g., where prostaglandins are involved) or treatment of autoimmunepathologies such as uveitis, type I diabetes, rheumatoid arthritis,chronic inflammatory disorders, inflammatory bowel diseases (e.g.,Crohn's disease and ulcerative colitis), multiple sclerosis, and indrug-eluting stents. Additional uses can include treatment of braindegenerative diseases, heart diseases, cancers, or hepatitis C. See, forexample, WO 2005/085295, WO 2004/010987, WO 03/097028, and WO2006/072562, each of which is incorporated by reference in its entirety.A class of S1P receptor agonists are described in provisional U.S.Application No. 60/956,111, filed Aug. 15, 2007, and PCT/US2008/073378,filed Aug. 15, 2008, each of which is incorporated by reference in itsentirety. See also provisional U.S. Application No. 61/231,539, filedAug. 5, 2009, and PCT/US2010/44607, filed Aug. 5, 2010, each of which isincorporated by reference in its entirety. See also provisional U.S.Application No. 61/440,254, filed Feb. 7, 2011, and PCT/US2012/23799filed Feb. 6, 2012, each of which is incorporated by reference in itsentirety.

Additional potential uses of an S1P modulating agent include, but arenot limited to, prevention or treatment of a pathological condition orsymptom in a mammal. For example, the condition can include inhibitedcell migration of oligodendrocyte precursor cells (OPCs).

Potential uses of an S1P receptor antagonist, and S1P4 receptor typeselective antagonists particularly, include, but are not limited to,prevention or treatment of a pathological condition or symptom in amammal.

LPA has been shown to be involved in lymphocyte trafficking and helpspromote entry of lymphocytes into secondary lymphoid organs (see Kanda,et al., Nat. Immunology (2008), 9:415-423). Therefore, the disclosedcompounds are expected to be useful for altering lymphocyte traffickingas a method for prolonging allograft survival, for exampletransplantation including solid organ transplants, treatment of graftvs. host disease, bone marrow transplantation, and the like.

An “ATX modulating agent” refers a compound or composition that iscapable of inducing a detectable change in ATX activity in vivo or invitro (e.g., at least 10% increase or decrease in ATX activity asmeasured by a given assay such as the assays described in the examplesand known in the art. A compound of the invention be an ATX modulatingagent, i.e., it can modulate the activity of ATX. For example, acompound of the invention can be an ATX inhibitor. The compound can be aselective ATX modulating agent. Being selective can mean that thecompound binds to ATX preferentially when exposed to a variety ofpotential binding partners. The compound can have a greater affinity forthe ATX, by at by at least 100-fold, by at least 50-fold, by at least10-fold, by at least 5-fold or by at least 2-fold, than for otherbinding partners. Affinity can be measured, for example, as adissociation constant (K_(d)), as an inhibition constant (such as IC₅₀),or another measure; provided that affinity is measured in a consistentfashion between ATX and the other binding partners it is compared to.

An inhibitor of ATX mediated activity can block interaction of ATX withits native substrate(s), such as LPC. For example, the inhibitor canshow an IC₅₀ value of less than 1 μM, less than 750 nM, less than 500nM, less than 250 nM, less than 100 nM, less than 50 nM, less than 25nM, or less than 10 nM, when measured in a FRET-based assay using FS-3substrate (see, e.g., Ferguson, C. G., et al., Org Lett. 2006 May 11;8(10): 2023-2026, which is incorporated by reference in its entirety).

Some substrates and inhibitors of ATX are described in WO 2011/151461,which is incorporated by reference in its entirety.

Potential uses of an ATX modulating agent include, but are not limitedto, prevention or treatment of a pathological condition or symptom in amammal. The pathological disorder can be an inflammatory disorder, anautoimmune disorder, a fibrosis of the lung, or a malignancy of thelung. Prevention or treatment of the pathological condition or symptomcan include administering to the mammal an effective amount of an ATXmodulating agent, e.g., an ATX inhibitor, to prevent, treat or reducesymptoms of the inflammatory disorder, autoimmune disorder, the fibrosisof the lung, or the malignancy of the lung. In one embodiment, theinflammatory disorder is rheumatoid arthritis (RA). In anotherembodiment, the autoimmune disorder is multiple sclerosis (MS). Aparticular example of lung fibrosis is an interstitial lung disease, forinstance, pulmonary fibrosis. See, for example, WO 2011/151461, which isincorporated by reference in its entirety.

In some embodiments, an ATX inhibitor of the present invention can beused to treat or prevent a demyelinating disease or disorder.Demyelinating diseases or disorders include multiple sclerosis,Guillain-Barre Syndrome, chronic inflammatory demyelinatingpolyneuropathy (CIDP), transverse myelitis, and optic neuritis, spinalcord injury, stroke or other ischemia, cerebral palsy,Charcot-Marie-Tooth disease (CMT), Sjogren-Larsson syndrome, Refsumdisease, Krabbe disease, Canavan disease, Alexander disease, nervedamage due to pernicious anemia, progressive multifocalleukoencephalopathy (PML), Lyme disease, tabes dorsalis due to untreatedsyphilis, demyelination due to exposure to an organophosphates,demyelination due to vitamin B12 deficiency or copper deficiency.

In addition, disclosed compounds can be useful as antagonists of thecannabinoid CB₁ receptor. CB₁ antagonism is associated with a decreasein body weight and an improvement in blood lipid profiles. The CB₁antagonism could be in concert with S1P receptor activity, or beindependent of activity at any S1P receptor.

In addition, disclosed compounds can be useful for inhibition of groupIVA cytosolic PLA₂ (cPLA₂). cPLA₂ catalyzes the release of eicosanoicacids (e.g., arachidonic acid). The eicosanoic acids are transformed topro-inflammatory eicosanoids such as prostaglandins and leukotrienes.Thus, disclosed compounds may be useful as anti-inflammatory agents.This inhibition could be in concert with S1P receptor activity, or beindependent of activity at any S1P receptor.

In addition, disclosed compounds may be useful for inhibition of themultiple substrate lipid kinase (MuLK). MuLK is highly expressed in manyhuman tumor cells and thus its inhibition might slow the growth orspread of tumors.

Neurological Disorders

MS can begin with a relapsing-remitting pattern of neurologicinvolvement, which then can progress to a chronic phase with increasingneurological damage. MS can be associated with the destruction ofmyelin, oligodendrocytes or axons localized to chronic lesions. Thedemyelination observed in MS may not always permanent and remyelinationhas been documented in early stages of the disease. Remyelination ofneurons can require oligodendrocytes.

The distal tip of an extending axon or neurite can include a specializedregion, known as the growth cone. Growth cones can sense the localenvironment and can guide axonal growth toward a neuron's target cell.Growth cones can respond to environmental cues, for example, surfaceadhesiveness, growth factors, neurotransmitters and electric fields. Thegrowth cones can advance at a rate of one to two millimeters per day.The growth cone can explore the area ahead of it and on either side, bymeans of elongations classified as lamellipodia and filopodia. When anelongation contacts an unfavorable surface, it can withdraw. When anelongation contacts a favorable growth surface, it can continue toextend and guides the growth cone in that direction. When the growthcone reaches an appropriate target cell a synaptic connection can becreated.

Nerve cell function can be influenced by contact between neurons andother cells in their immediate environment (Rutishauser, et al., 1988,Physiol. Rev. 68:819, which is incorporated by reference in itsentirety). These cells can include specialized glial cells,oligodendrocytes in the central nervous system (CNS), and Schwann cellsin the peripheral nervous system (PNS), which can sheathe the neuronalaxon with myelin (Lemke, 1992, in An Introduction to MolecularNeurobiology, Z. Hall, Ed., p. 281, Sinauer, each of which isincorporated by reference in its entirety). LPA causes the collapse ofthe neuron growth cone and tends to inhibit or reverse the morphologicaldifferentiation of many neuronal cell lines (see Gendaszewska-Darmach,Acta Biochimica Polonica (2008), 55(2):227-240). CNS neurons can havethe inherent potential to regenerate after injury, but they can beinhibited from doing so by inhibitory proteins present in myelin(Brittis et al., 2001, Neuron 30:11-14; Jones et al., 2002, J. Neurosci.22:2792-2803; Grimpe et al., 2002, J. Neurosci.: 22:3144-3160, each ofwhich is incorporated by reference in its entirety).

Several myelin inhibitory proteins found on oligodendrocytes have beencharacterized. Known examples of myelin inhibitory proteins can includeNogoA (Chen et al., Nature, 2000, 403, 434-439; Grandpre et al., Nature2000, 403, 439-444, each of which is incorporated by reference in itsentirety), myelin associated glycoprotein (MAG) (McKerracher et al.,1994, Neuron 13:805-811; Mukhopadhyay et al., 1994, Neuron 13:757-767,each of which is incorporated by reference in its entirety) oroligodendrocyte glycoprotein (OM-gp), Mikol et al., 1988, J. Cell. Biol.106:1273-1279, each of which is incorporated by reference in itsentirety). Each of these proteins can be a ligand for the neuronal Nogoreceptor-1 (NgR1 (Wang et al., Nature 2002, 417, 941-944; Grandpre etal., Nature 2000, 403, 439-444; Chen et al., Nature, 2000, 403, 434-439;Domeniconi et al., Neuron 2002, published online Jun. 28, 2002, each ofwhich is incorporated by reference in its entirety).

Nogo receptor-1 (NgR1) is a GPI-anchored membrane protein that contains8 leucine rich repeats (Fournier et al., 2001, Nature 409:341-346, whichis incorporated by reference in its entirety). Upon interaction withinhibitory proteins (e.g., NogoA, MAG and OM-gp), the NgR1 complex cantransduce signals that lead to growth cone collapse and inhibition ofneurite outgrowth.

There is a need for molecules and methods for inhibiting NgR1-mediatedgrowth cone collapse and the resulting inhibition of neurite outgrowth.Additionally, there is a need for molecules which increase neuronalsurvival and axon regeneration, particularly for the treatment ofdisease, disorders or injuries that involve axonal injury, neuronal oroligodendrocyte cell death, demyelination or dymyelination or generallyrelate to the nervous system.

Such diseases, disorders or injuries can include, but are not limitedto, multiple sclerosis (MS), progressive multifocal leukoencephalopathy(PML), encephalomyelitis (EPL), central pontine myelolysis (CPM),adrenoleukodystrophy, Alexander's disease, Pelizaeus Merzbacher disease(PMZ), Globoid cell Leucodystrophy (Krabbe's disease) and WallerianDegeneration, optic neuritis, transverse myelitis, amylotrophic lateralsclerosis (ALS), Huntington's disease, Alzheimer's disease, Parkinson'sdisease, spinal cord injury, traumatic brain injury, post radiationinjury, neurologic complications of chemotherapy, stroke, acute ischemicoptic neuropathy, vitamin E deficiency, isolated vitamin E deficiencysyndrome, AR, Bassen-Kornzweig syndrome, Marchiafava-Bignami syndrome,metachromatic leukodystrophy, trigeminal neuralgia, or Bell's palsy.Among these diseases, MS may the most widespread, affectingapproximately 2.5 million people worldwide.

Various disease-modifying treatments may be available for MS, includingthe use of corticosteroids and immunomodulating agents such asinterferon beta or Tysabri®. In addition, because of the central role ofoligodendrocytes and myelination in MS, there have been efforts todevelop therapies to increase oligodendrocyte numbers or enhancemyelination. See, e.g., Cohen et al., U.S. Pat. No. 5,574,009; Chang etal., N. Engl. J. Med. 346: 165-73 (2002), each of which is incorporatedby reference in its entirety. However, there remains an urgent need todevise additional therapies for MS and other demyelination anddismyelination disorders.

A compound of the invention, or a pharmaceutically acceptable saltthereof, can promote myelination or remyelination. A method can includeadministering a compound of the invention, or a pharmaceuticallyacceptable salt thereof, to cells. A method of promoting oligodendrocyteprogenitor cell differentiation can include administering a compound ofthe invention, or a pharmaceutically acceptable salt thereof, to cells.A method of treating multiple sclerosis can include administering acompound of the invention, or a pharmaceutically acceptable saltthereof, to a subject.

A number of studies have shown that ATX is expressed in non-pathologicalconditions, throughout development, with high expression levels in theCNS among other tissues. ATX mRNA was identified as highly upregulatedduring oligodendrocyte differentiation and ATX protein expression isalso apparent in maturing ODCs, temporally correlated with the processof myelination. Finally, in the adult brain ATX is expressed insecretory epithelial cells, such as the choroid plexus, ciliary, irispigment, and retinal pigment epithelial cells, whereas there is evidencefor ATX expression in leptomeningeal cells and cells of the CNSvasculature. See, for example, Fuss, B., et al., J Neurosci 17,9095-9103 (1997); Kawagoe, H., et al. Genomics 30, 380-384 (1995); Lee,H. Y., et al. J Biol Chem 271, 24408-24412 (1996); Narita, M., et al., JBiol Chem 269, 28235-28242 (1994); Bachner, D., et al., Mechanisms ofDevelopment 84, 121-125 (1999); Awatramani, R., et al., Nat Genet 35,70-75 (2003); Li, Y., et al., J Neurol Sci 193, 137-146 (2002); Dugas,J. C., et al., J Neurosci 26, 10967-10983 (2006); Fox, M. A., et al.,Molecular and Cellular Neuroscience 27, 140-150 (2004); Hoelzinger, D.B., et al., Neoplasia 7, 7-16 (2005); and Sato, K., et al., J Neurochem92, 904-914 (2005); each of which is incorporated by reference in itsentirety.

Although neurons and astrocytes do not seem to express ATX underphysiological conditions, ATX is highly upregulated in astrocytesfollowing brain lesion. Two hallmarks of reactive astrogliosis can beinduced by LPA itself: hypertrophy of astrocytes and stress fiberformation. This may indicate an autoregulation loop of astrocyticactivation, in which astrocytes upregulate the LPA-generating enzyme ATXand become activated by its metabolite LPA, while increased amounts ofthe metabolite inhibit the catalytic activity of ATX. See, e.g.,Savaskan, N. E., et al., Cell Mol Life Sci 64, 230-243 (2007); Ramakers,G. J, & Moolenaar, W. H., Exp Cell Res 245, 252-262 (1998); and vanMeeteren, L. A., et al., J Biol Chem 280, 21155-21161 (2005); each ofwhich is incorporated by reference in its entirety.

ATX expression levels were shown to be elevated in glioblastomamultiform samples, and ATX was shown to augment invasiveness of cellstransformed with ras, a key signaling molecule that promotesgliomagenesis. ATX expression was also detected in primary tumor tissuesfrom neuroblastoma patients and retinoic acid induced expression of ATXin N-myc-amplified neuroblastoma cells.

There is significant evidence for ATX signaling in demyelinationprocesses and in other neurodegenerative conditions. As noted above, ithas been reported that addition of LPA to dorsal root fibers in ex vivoculture causes demyelination, whereas LPC fails to cause significantdemyelination of nerve fibers in ex vivo cultures without furtheraddition of recombinant ATX to the culture. Addition of recombinant ATXcaused significant demyelination at equivalent levels to LPA presumabledue to conversion of LPC to LPA through the enzymatic activity of ATX.In addition, injury induced demyelination was attenuated by about 50% inatx^(+/−) mice over their wild type counterparts (Nagai, et al.,Molecular Pain (2010), 6:78).

ATX protein levels were found deregulated in an animal model of MS(experimental autoimmune encephalitis; EAE) at the onset of clinicalsymptoms. See, e.g., Hoelzinger, D. B., et al. Neoplasia 7, 7-16 (2005);Nam, S. W., et al., Oncogene 19, 241-247 (2000); Kawagoe, H., et al.,Cancer Res 57, 2516-2521 (1997); Dufner-Beattie, J., et al., MolCarcinog 30, 181-189 (2001); Umemura, K., et al., Neuroscience Letters400, 97-100 (2006); and Fuss, B., et al., J Neurosci 17, 9095-9103(1997); each of which is incorporated by reference in its entirety.Moreover, significant ATX expression was been detected in thecerebrospinal fluid of patients suffering with multiple sclerosis (MS),while completely lacking from the control samples, suggesting a role forATX in maintenance of cerebrospinal fluid homeostasis duringpathological/demyelinating conditions. Hammack, B. N., et al. Proteomicanalysis of multiple sclerosis cerebrospinal fluid. Mult Scler 10,245-260 (2004); and Dennis, J., et al., J Neurosci Res 82, 737-742(2005); each of which is incorporated by reference in its entirety.

Interestingly, ATX mRNA expression was found to be elevated in thefrontal cortex of Alzheimer-type dementia patients indicating apotential involvement for ATX signaling in neurodegenerative diseases.LPA receptors are enriched in the CNS and their expression patternssuggest their potential involvement in developmental process includingneurogenesis, neuronal migration, axon extension and myelination.Noteworthy, only two receptors have the same spatiotemporal expressionas ATX in the CNS (Contos, J. J., et al., Mol Cell Biol 22, 6921-6929(2002); Jaillard, C, ei al, Edg8/S1 P5: an oligodendroglial receptorwith dual function on process retraction and cell survival. J Neurosci25, 1459-1469 (2005); and Saba, J. D. Journal of cellular biochemistry92, 967-992 (2004); each of which is incorporated by reference in itsentirety). LPAi and S1P5 are specific for ODCs, and their expressionhighly correlates with the process of myelination. LPA1 is expressed inrestricted fashion within the neuroblasts of the neuroproliferatveVentricular Zone (VZ) of the developing cortex, in the dorsal olfactorybulb, along the pial cells of neural crest origin, and in developingfacial bone tissue. Expression is observed during E11-E18, correspondingto a time period during which neurogenesis occurs. LPA1 expression isundetectable in the VZ after this point, to reappear during the firstpostnatal week within ODCs. Notably, Schwann cells (the myelinatingcells of the Peripheral Nervous System; PNS) express high levels of LPA1early in development and persistently throughout life, suggesting aninfluence of LPA on myelinating processes (Weiner. J. A. & Chun, J.,Proc Natl Acad Sci USA 96, 5233-5238 (1999), which is incorporated byreference in its entirety).

The above data strongly support a critical role for ATX and LPAsignaling in neuronal development, oligodendrocyte differentiation andmyelination, as well as possibly in the autoregulation of astrocyteactivation. Moreover, the regulation of ATX and thus LPA production atlocal sites of CNS injury, inflammatory or autoimmune, could contributeto tissue homeostasis through the numerous effects of LPA. Asdemyelination and deregulated cerebrospinal fluid homeostasis are thehallmarks of multiple sclerosis, a role of ATX and LPA signaling in thepathophysiology of multiple sclerosis seems very likely.

The S1P modulating agents and/or ATX modulating agents of the inventioncan be used to various forms of MS including relapsing-remitting,secondary-progressive, primary-progressive, and progressive-relapsingforms. In addition, S1P modulating agents and/or ATX modulating agentsof the invention can be used alone or in conjunction with other agentsto treat or prevent MS. In a preferred embodiment, the compounds of theinvention can be used to treat or prevent MS in combination with animmunomodulating therapy such as corticosteroids, beta interferon-1a(such as Avonex® or Rebif®), beta interferon-1b (Betaseron®),natalizumab (Tysabri®), glatiramer, and mitoxantrone.

Pain Mediation

Pain experienced by mammals can be divided into two main categories:acute pain (or nociceptive) and chronic pain which can be subdividedinto chronic inflammatory pain and chronic neuropathic pain. Acute painis a response to stimulus that causes tissue injury and is a signal tomove away from the stimulus to minimize tissue damage. Chronic pain, onthe other hand, serves no biological function and develops as a resultof inflammation caused by tissue damage (inflammatory pain) or by damageto the nervous system such as demyelination (neuropathic pain). Chronicpain is generally characterized by stimulus-independent, persistent painor by abnormal pain perception triggered by innocuous stimuli.

LPA has been found to be a mediator of both inflammatory pain andneuropathic pain. The transient receptor potential channel TRPV1 isknown to be the originator of inflammatory pain. LPA has been shown todirectly activate TRPV1 thereby creating pain stimulus by binding to itsintracellular C-terminus (Tigyi, Nature Chemical Biology (January 2012),8:22-23).

LPA has also been shown to play a role in neuropathic pain. For example,sciatic nerve injury has been shown to induce demyelination,down-regulation of myelin-associated glycoprotein (MAG) and damage toSchwann cell partitioning of C-fiber-containing Remak bundles in thesciatic nerve and dorsal root. However, demyelination, MAGdown-regulation and Remak bundle damage in the dorsal root wereabolished in LPA₁ receptor-deficient (Lpar1^(−/−)) mice (Nagai, et al.,Molecular Pain (2010), 6:78).

Thus compounds of the invention are useful in treating or preventingchronic pain such as inflammatory pain and neuropathic pain in mammals.

Rheumatoid Arthritis (RA)

Studies in human and animal models of RA suggest that ATX plays a rolein the development and progress of the disease. For example, increasedATX mRNA expression was detected in synovial fibroblasts (SFs) fromanimal models of RA during differential expression profiling, and humanRA SFs were shown to express mRNA for both ATX and LPARs (Aidinis, V.,et al., PLoS genetics 1, e48 (2005); Zhao, C, et al., Molecularpharmacology 73, 587-600 (2008); each of which is incorporated byreference in its entirety). ATX is overexpressed from activated SFs inarthritic joints, both in animal models and human patients (see WO2011/151461). ATX expression was shown to be induced from TNF, the majorpro-inflammatory factor driving RA.

Disease development was assessed in well-established animal models ofRA. When ATX expression was conditionally ablated specifically in SFs,the lack of ATX expression in the joints resulted in marked decreasedinflammation and synovial hyperplasia. This suggested an activeinvolvement of the ATX-LPA axis in the pathogenesis of the disease.Similar results were also obtained with pharmacologic inhibition of ATXenzymatic activity and LPA signaling. A series of ex vivo experiments onprimary SFs revealed that ATX, through LPA production, stimulatesrearrangements of the actin cytoskeleton, proliferation and migration tothe extracellular matrix (ECM), as well as the secretion ofproinflammatory cytokines and matrix metalloproteinases (MMPs).Moreover, the LPA effect was shown to be synergistic with TNF anddependent on the activation of MAPK cellular signaling pathways. See,e.g., Armaka, M., et al., The Journal of experimental medicine 205,331-337 (2008); which is incorporated by reference in its entirety.

In one embodiment a method for treating an individual with RA or theindividual at risk of suffering thereof comprises administering to saidindividual an S1P modulating agent and/or ATX modulating agent of theinvention in conjunction with an anti-TNF antibody for use in thetreatment of RA. Examples of suitable anti-TNF antibodies areadalimumab, etanercept, golimumab, and infliximab (Taylor P C, FeldmannM. Anti-TNF biologic agents: still the therapy of choice for rheumatoidarthritis. Nat Rev Rheumatol. 2009 October; 5(10):578-82).

Pulmonary Fibrosis

Evidence also suggests a role for ATX in pulmonary fibrosis. Micelacking lysophosphatidic acid (LPA) receptor 1 (LPAR1) were protectedfrom Bleomycin (BLM)-induced pulmonary fibrosis and mortality,suggesting a major role for LPA in disease pathophysiology. The majorityof circulating LPA is produced by the phospholipase D activity ofAutotaxin (ATX) and the hydrolysis of lysophosphatidylcholine (LPC).Increased ATX expression has been previously reported in thehyperplastic epithelium of fibrotic lungs of human patients and animalmodels.

Therefore, we hypothesized that genetic or pharmacologic inhibition ofATX activity would reduce local or circulating LPA levels and henceattenuate disease pathogenesis.

Lung Cancer

Increased ATX expression has been detected in a large number ofmalignancies, including mammary, thyroid, hepatocellular and renal cellcarcinomas, glioblastoma and neuroblastoma, as well as NSCLC.Strikingly, transgenic overexpression of ATX was shown to inducespontaneous mammary carcinogenesis. In accordance, in vitro ATXoverexpression in various cell types promotes proliferation andmetastasis while inhibiting apoptosis. LPA's actions are concordant withmany of the “hallmarks of cancer”, indicating a role for LPA in theinitiation or progression of malignant disease. Indeed LPA levels aresignificantly increased in malignant effusions, and its receptors areaberrantly expressed in several human cancers.

See, for example: Euer, N., et al., Anticancer Res 22, 733-740 (2002);Liu, S., et al., Cancer Cell 15, 539-550 (2009); Zhang, G., et al., ChinMed J (Engl) 112, 330-332 (1999); Stassar, M. J., et al., Br J Cancer85. 1372-1382 (2001); Kishi, Y., et al., J Biol Chem 281, 17492-17500(2006); Kawagoe, H., et al., Cancer Res 57, 2516-2521 (1997); Yang, Y.,et al., Am J Respir Cell Mol Biol 21, 216-222 (1999); and Toews, M. L.,et al. Biochim Biophys Acta 1582, 240-250 (2002); each of which isincorporated by reference in its entirety.

In cases where a compound of the invention can be sufficiently basic oracidic to form stable nontoxic acid or base salts, preparation andadministration of the compounds as pharmaceutically acceptable salts maybe appropriate. Examples of pharmaceutically acceptable salts can beorganic acid addition salts formed with acids which form a physiologicalacceptable anion, for example, tosylate, methanesulfonate, acetate,citrate, malonate, tartarate, succinate, benzoate, ascorbate,α-ketoglutarate, or α-glycerophosphate. Inorganic salts may also beformed, including hydrochloride, sulfate, nitrate, bicarbonate, andcarbonate salts.

Pharmaceutically acceptable salts may be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid affording aphysiologically acceptable anion. Alkali metal (for example, sodium,potassium or lithium) or alkaline earth metal (for example calcium)salts of carboxylic acids can also be made.

Pharmaceutically-acceptable base addition salts can be prepared frominorganic and organic bases. Salts from inorganic bases, can include butare not limited to, sodium, potassium, lithium, ammonium, calcium ormagnesium salts. Salts derived from organic bases can include, but arenot limited to, salts of primary, secondary or tertiary amines, such asalkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines,di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenylamines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines,di(substituted alkenyl) amines, tri(substituted alkenyl) amines,cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl)amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines,disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines,aryl amines, diaryl amines, triaryl amines, heteroaryl amines,diheteroaryl amines, triheteroaryl amines, heterocyclic amines,diheterocyclic amines, triheterocyclic amines, or mixed di- andtri-amines where at least two of the substituents on the amine can bedifferent and can be alkyl, substituted alkyl, alkenyl, substitutedalkenyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, or heterocyclic and the like. Alsoincluded can be amines where the two or three substituents, togetherwith the amino nitrogen, form a heterocyclic or heteroaryl group.Non-limiting examples of amines can include, isopropylamine, trimethylamine, diethyl amine, tri(iso-propyl) amine, tri(n-propyl) amine,ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine,histidine, caffeine, procaine, hydrabamine, choline, betaine,ethylenediamine, glucosamine, N-alkylglucamines, theobromine, purines,piperazine, piperidine, morpholine, or N-ethylpiperidine, and the like.Other carboxylic acid derivatives can be useful, for example, carboxylicacid amides, including carboxamides, lower alkyl carboxamides, ordialkyl carboxamides, and the like.

Pharmaceutical compositions can include a compound of the invention, ora pharmaceutically acceptable salt thereof. More particularly, suchcompounds can be formulated as pharmaceutical compositions usingstandard pharmaceutically acceptable carriers, fillers, solubilizingagents and stabilizers known to those skilled in the art. For example, apharmaceutical composition including a compound of the invention, or asalt, analog, derivative, or modification thereof, as described herein,is used to administer the appropriate compound to a subject.

The compounds of the invention, or a pharmaceutically acceptable saltthereof, are useful for treating a disease or disorder associated withS1P receptor mediated activity, and/or ATX activity. In one embodiment,a therapeutically effective amount of a compound of the invention, or apharmaceutically acceptable salt thereof, is administered to a subjectin need thereof. In another embodiment, a pharmaceutical compositioncomprising a therapeutically effective amount of a compound of theinvention, or a pharmaceutically acceptable salt thereof, and apharmaceutically-acceptable carrier is administered to a subject in needthereof.

The compounds of the invention can be used in combination with at leastone further active ingredient, such as a medicament used in thetreatment of multiple sclerosis such as Tysabri®, dimethyl fumarate, aninterferon (such as pegylated or non-pegylated interferons, preferablyinterferon β-1a or pegylated interferon β-1a), glatiramer acetate, acompound improving vascular function, an immunomodulating agent (such asFingolimod, cyclosporins, rapamycins or ascomycins, or theirimmunosuppressive analogs, e.g. cyclosporine A, cyclosporine G, FK-506,ABT-281, ASM981, rapamycin, 40-O-(2-hydroxyl)ethyl-rapamycin etc.);corticosteroids; cyclophosphamide; azathioprine; mitoxanthrone,methotrexate; leflunomide; mizoribine; mycophenolic add; mycophenolatemofetil; 15-deoxyspergualine; diflucortolone valerate; difluprednate;Alclometasone dipropionate; amcinonide; amsacrine; asparaginase;azathioprine; basiliximab; beclometasone dipropionate; betamethasone;betamethasone dipropionate; betamethasone phosphate sodique;betamethasone valerate; budesonide; captopril; chlormethinechlorhydrate; clobetasol propionate; cortisone acetate; cortivazol;cyclophosphamide; cytarabine; daclizumab; dactinomycine; desonide;desoximetasone; dexamethasone; dexamethasone acetate; dexamethasoneisonicotinate; dexamethasone metasulfobenzoate sodique;dexamethasonephosphate; dexamethasone tebutate; dichlorisone acetate;doxorubicinee chlorhydrate; epirubicine chlorhydrate; flucloroloneacetonide; fludrocortisone acetate; fludroxycortide; flumetasonepivalate; flunisolide; fluocinolone acetonide; fluocinonide;fluocortolone; fluocortolone hexanoate; fluocortolone pivalate;fluorometholone; fluprednidene acetate; fluticasone propionate;gemcitabine chlorhydrate; halcinonide; hydrocortisone; hydrocortisoneacetate; hydrocortisone butyrate; hydrocortisone hemisuccinate;melphalan; meprednisone; mercaptopurine; methylprednisolone;methylprednisolone acetate; methylprednisolone hemisuccinate;misoprostol; muromonab-cd3; mycophenolate mofetil; paramethansoneacetate; prednazoline, prednisolone; prednisolone acetate; prednisolonecaproate; prednisolone metasulfobenzoate sodique; prednisolone phosphatesodique; prednisone; prednylidene; rifampicine; rifampicine sodique;tacrolimus; teriflunomide; thalidomide; thiotepa; tixocortol pivalate;triamcinolone; triamcinolone acetonide hemisuccinate; triamcinolonebenetonide; triamcinolone diacetate; triamcinolone hexacetonide;immunosuppressive monoclonal antibodies, e.g., monoclonal antibodies toleukocyte receptors, e.g., MHC, CD2, CD3, CD4, CD7, CD20 (e.g.,rituximab and ocrelizumab), CD25, CD28, B7, CD40, CD45, CD56 (e.g.,daclizumab), or CD58 or their ligands; or other immunomodulating agentycompounds, e.g. CTLA41g, or other adhesion molecule inhibitors, e.g.mAbs or low molecular weight inhibitors including Selectin antagonistsand VLA-4 antagonists (such as Tysabri®); remyelinating agents such asBIIB033. Compounds of the invention can also be used in combination withagents which treat the symptoms of multiple sclerosis such asfampridine.

The dose of a compound of the invention, or a pharmaceuticallyacceptable salt thereof, administered to a subject can be less than 10μg, less than 25 μg, less than 50 μg, less than 75 μg, less than 0.10mg, less than 0.25 mg, less than 0.5 mg, less than 1 mg, less than 2.5mg, less than 5 mg, less than 10 mg, less than 15 mg, less than 20 mg,less than 50 mg, less than 75 mg, less than 100 mg, or less than 500 mg.

Administering can include administering by topical, enteral, parenteral,transdermal, transmucosal, inhalational, intracisternal, epidural,intravaginal, intravenous, intramuscular, subcutaneous, intradermal orintravitreal administration.

The duration of administering can be less than 30 seconds, less than 1minute, about 1 minute, between 1 minute and 5 minutes, between 5minutes and 10 minutes, between 10 minutes and 20 minutes, between 20minutes and 30 minutes, between 30 minutes and 1 hour, between 1 hourand 3 hours, between 3 hours and 6 hours, between 6 hours and 12 hours,between 12 hours and 24 hours or for more than 24 hours.

Administering the inhibitor or compound can include multipleadministrations. The duration between administrations can be less than30 seconds, less than 1 minute, about 1 minute, between 1 minute and 5minutes, between 5 minutes and 10 minutes, between 10 minutes and 20minutes, between 20 minutes and 30 minutes, between 30 minutes and 1hour, between 1 hour and 3 hours, between 3 hours and 6 hours, between 6hours and 12 hours, between 12 hours and 24 hours or for more than 24hours.

The duration between successive administrations can be less than 30seconds, less than 1 minute, about 1 minute, between 1 minute and 5minutes, between 5 minutes and 10 minutes, between 10 minutes and 20minutes, between 20 minutes and 30 minutes, between 30 minutes and 1hour, between 1 hour and 3 hours, between 3 hours and 6 hours, between 6hours and 12 hours, between 12 hours and 24 hours, between 24 hours and48 hours, between 48 hours and 72 hours, between 72 hours and 1 week orbetween 1 week and 2 weeks.

Administering an inhibitor or compound to cells can include cells of anin vitro or in vivo system or model. The cells can be part of a cellline. The cell line can be a primary or secondary cell line. The cellline can be an immortal cell line. The cells can be ruptured and be inthe form of a cell lysate. The cells can be part of a living organism,i.e., a subject, for example, a mammal. A mammal can include a rat, amouse, a gerbil, a hamster, a rabbit or a human. The human can be asubject or a patient.

A method can further include monitoring a property of a sample or asubject. A sample can be removed from a subject. For instance, a samplecan include a sample of cells or a tissue from a subject. A sample caninclude blood, plasma, or neuronal tissue including neurons or glialcells. A sample can also remain in the subject. For example, a samplecan be a tissue or cells that are observed within the patient.

A method can further include providing untreated control cells, sampleor subject and measuring a property of a sample of the untreated controlcells, sample or subject.

A property can include the presence or absence of a molecule, theconcentration of a molecule, for example myelin basic protein, myelinassociated glycoprotein or myelin oligodendrocyte glycoprotein. In someembodiments, determining the presence of a molecule can includedetermining the concentration of the molecule, determining the purity ofthe molecule or determining the quantity of the molecule.

A property can be the conductivity of a tissue or cell. A property canbe an emission, for example, electromagnetic radiation.

Monitoring a property can include observing the property of the sampleor subject alone. Monitoring a property can include monitoring theproperty before the sample or subject has been administered a compoundof the invention. Monitoring a property can include monitoring theproperty after the sample or subject has been administered a compound.Monitoring a property can include monitoring a property after the sampleor subject has been administered a known concentration of a compound.

Monitoring a property of a sample or subject can include observing theproperty through a microscope. Monitoring a property of the compositioncan include measuring the property using a microscope. Monitoring aproperty of the composition can include monitoring the property usingstill photography or movies. The photography or movies can be on filmmedia or digital form. Monitoring a property can include taking a scan,for example, an MRI or CT scan.

Promoting myelination, remyelination or oligodendrocyte progenitor celldifferentiation can prevent or can treat a pathological condition orsymptom in a mammal. A number of diseases or disorders involvedemyelination of the central or peripheral nervous system which canoccur for a number of reasons such as immune dysfunction as in multiplesclerosis, encephalomyelitis, Guillain-Barre Syndrome, chronicinflammatory demyelinating polyneuropathy (CIDP), transverse myelitis,and optic neuritis; demyelination due to injury such as spinal cordinjury, traumatic brain injury, stroke, acute ischemic optic neuropathy,or other ischemia, cerebral palsy, neuropathy (e.g. neuropathy due todiabetes, chronic renal failure, hypothyroidism, liver failure, orcompression of the nerve), post radiation injury, and central pontinemyelolysis (CPM); inherited conditions such as Charcot-Marie-Toothdisease (CMT), Sjogren-Larsson syndrome, Refsum disease, Krabbe disease,Canavan disease, Alexander disease, Friedreich's ataxia,Pelizaeus-Merzbacher disease, Bassen-Kornzweig syndrome, metachromaticleukodystrophy (MLD), adrenoleukodystrophy, and nerve damage due topernicious anemia; viral infection such as progressive multifocalleukoencephalopathy (PML), Lyme disease, or tabes dorsalis due tountreated syphilis; toxic exposure due to chronic alcoholism (which is apossible cause of Marchiafava-Bignami disease), chemotherapy, orexposure to chemicals such as organophosphates; or dietary deficienciessuch as vitamin B12 deficiency, vitamin E deficiency, and copperdeficiency. Some demyelination disorders can have unknown or multiplecauses such as trigeminal neuralgia, Marchiafava-Bignami disease andBell's palsy. In addition, demyelination can contribute to neuropathicpain. Compounds of the invention are expected to be useful in treatingdemyelination disorders.

Since LPA is a proinflammatory factor reducing the amount of LPAproducted by inhibiting ATX is useful for treating inflammatorydisorders such as asthma, allergies, arthritis, inflammatoryneuropathies, transplantation rejection, Crohn's disease, ulcerativecolitis, lupus erythematosis, psoriasis, an inflammatory bowelcondition, and diabetes.

LPA has been shown to be involved in wound healing and stimulates theproliferation and migration of endothelial cells promoting processessuch as angiogenesis. However, these same processes when deregulated canpromote tumor growth and metastasis, and LPA is thought to contribute tothe development, progression, and metastasis of several types of cancerincluding ovarian, prostate, melanoma, breast, head and neck cancers(see Gendaszewska-Darmach, Acta Biochimica Polonica (2008),55(2):227-240). In addition, since ATX is located outside the cell incirculation, ATX inhibitors are expected to be of most benefit outsidethe cell. Therefore, ATX inhibitors are expected to be useful intreating cancer, particularly multidrug resistant (MDR) cancers wheredrug efflux mechanisms are the largest contributor to the drugresistance.

A compound of the invention, or a pharmaceutically acceptable saltthereof, formulated as a pharmaceutical composition and administered toa mammalian host, such as a human patient in a variety of forms adaptedto the chosen route of administration, e.g., orally or parenterally, aseyedrops, by intravenous, intramuscular, topical or subcutaneous routes.In addition, the term “administer” or “administering” encompassesdelivering a compound of the invention as a prodrug which is convertedor metabolized in the body of the mammal into a compound of theinvention. In one embodiment, a compound of the invention isadministered in a non-prodrug form. In another embodiment, the compoundis administered as a prodrug which is metabolized to a compound of theinvention in the body of a mammal.

Thus, a compound of the invention, or a pharmaceutically acceptable saltthereof, may be systemically administered, e.g., orally, in combinationwith a pharmaceutically acceptable vehicle such as an inert diluent oran assimilable edible carrier. They may be enclosed in hard or softshell gelatin capsules, may be compressed into tablets, or may beincorporated directly with the food of the patient's diet. For oraltherapeutic administration, the active compound may be combined with oneor more excipients and used in the form of ingestible tablets, buccaltablets, troches, capsules, elixirs, suspensions, syrups, or wafers, andthe like. Such compositions and preparations should contain at leastabout 0.1% of active compound. The percentage of the compositions andpreparations may, of course, be varied and may conveniently be betweenabout 2 to about 60% of the weight of a given unit dosage form. Theamount of active compound in such therapeutically useful compositionscan be such that an effective dosage level will be obtained.

The tablets, troches, pills, capsules, and the like can include thefollowing: binders such as gum tragacanth, acacia, corn starch orgelatin; excipients such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch, alginic acid and the like; alubricant such as magnesium stearate; or a sweetening agent such assucrose, fructose, lactose or aspartame or a flavoring agent such aspeppermint, oil of wintergreen, or cherry flavoring may be added. Whenthe unit dosage form is a capsule, it may contain, in addition tomaterials of the above type, a liquid carrier, such as a vegetable oilor a polyethylene glycol. Various other materials may be present ascoatings or to otherwise modify the physical form of the solid unitdosage form. For instance, tablets, pills, or capsules may be coatedwith gelatin, wax, shellac or sugar and the like. A syrup or elixir maycontain the active compound, sucrose or fructose as a sweetening agent,methyl or propylparabens as preservatives, a dye and flavoring such ascherry or orange flavor. Of course, any material used in preparing anyunit dosage form should be pharmaceutically acceptable and substantiallynon-toxic in the amounts employed. In addition, the active compound maybe incorporated into sustained-release preparations and devices.

The active compound may also be administered intravenously orintraperitoneally by infusion or injection. Solutions of the activecompound or its salts can be prepared in water, optionally mixed with anontoxic surfactant. Dispersions can also be prepared in glycerol,liquid polyethylene glycols, triacetin, and mixtures thereof and inoils. Under ordinary conditions of storage and use, these preparationscan contain a preservative to prevent the growth of microorganisms.

Exemplary pharmaceutical dosage forms for injection or infusion caninclude sterile aqueous solutions or dispersions or sterile powderscomprising the active ingredient which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. In all cases, theultimate dosage form should be sterile, fluid and stable under theconditions of manufacture and storage. The liquid carrier or vehicle canbe a solvent or liquid dispersion medium comprising, for example, water,ethanol, a polyol (for example, glycerol, propylene glycol, liquidpolyethylene glycols, and the like), vegetable oils, or nontoxicglyceryl esters, and mixtures thereof. The proper fluidity can bemaintained, for example, by the formation of liposomes, by themaintenance of the required particle size in the case of dispersions orby the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, or thimerosal, and the like. In many cases, it will be preferableto include isotonic agents, for example, sugars, buffers or sodiumchloride. Prolonged absorption of the injectable compositions can bebrought about by the use in the compositions of agents delayingabsorption, for example, aluminum monostearate or gelatin.

Sterile injectable solutions can be prepared by incorporating the activecompound in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfilter sterilization. In the case of sterile powders for the preparationof sterile injectable solutions, the preferred methods of preparationcan be vacuum drying and the freeze drying techniques, which can yield apowder of the active ingredient plus any additional desired ingredientpresent in the previously sterile-filtered solutions.

For topical administration, a compound of the invention may be appliedin pure form, e.g., when they are liquids. However, it can be generallybe desirable to administer them to the skin as compositions orformulations, in combination with a dermatologically acceptable carrier,which may be a solid or a liquid.

Exemplary solid carriers can include finely divided solids such as talc,clay, microcrystalline cellulose, silica, alumina and the like. Usefulliquid carriers include water, alcohols or glycols orwater-alcohol/glycol blends, in which the present compounds can bedissolved or dispersed at effective levels, optionally with the aid ofnon-toxic surfactants. Adjuvants such as fragrances and additionalantimicrobial agents can be added to optimize the properties for a givenuse. The resultant liquid compositions can be applied from absorbentpads, used to impregnate bandages and other dressings, or sprayed ontothe affected area using pump-type or aerosol sprayers.

Thickeners such as synthetic polymers, fatty acids, fatty acid salts oresters, fatty alcohols, modified celluloses or modified mineralmaterials can also be employed with liquid carriers to form spreadablepastes, gels, ointments, soaps, and the like, for application directlyto the skin of the user.

Examples of useful dermatological compositions which can be used todeliver a compound of the invention to the skin are known to the art;for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S.Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman(U.S. Pat. No. 4,820,508), each of which is incorporated by reference inits entirety.

Useful dosages of the compounds of the invention can be determined bycomparing their in vitro activity, and in vivo activity in animalmodels. Methods for the extrapolation of effective dosages in mice, andother animals, to humans are known to the art; for example, see U.S.Pat. No. 4,938,949, which is incorporated by reference in its entirety.

Generally, the concentration of a compound(s) of the invention in aliquid composition, such as a lotion, can be from about 0.1 to about 25weight percent, preferably from about 0.5-10 weight percent. Theconcentration in a semi-solid or solid composition such as a gel or apowder can be about 0.1-5 wt-%, preferably about 0.5-2.5 weight percentbased on the total weight of the composition.

The amount of the compound, or an active salt or derivative thereof,required for use in treatment can vary not only with the particular saltselected but also with the route of administration, the nature of thecondition being treated and the age and condition of the patient and canbe ultimately at the discretion of the attendant physician or clinician.In general, however, a dose can be in the range of from about 0.1 toabout 10 mg/kg of body weight per day.

The compound can be conveniently administered in unit dosage form; forexample, containing 0.01 to 10 mg, or 0.05 to 1 mg, of active ingredientper unit dosage form. In some embodiments, a dose of 5 mg/kg or less canbe suitable.

The active ingredient can be administered so as to achieve a desiredpeak plasma concentration of the active compound. The desired peakplasma concentration can be from about 0.5 μM to about 75 μM,preferably, about 1 μM to 50 μM, or about 2 μM to about 30 μM. This maybe achieved, for example, by the intravenous injection of a 0.05 to 5%solution of the active ingredient, optionally in saline, or orallyadministered as a bolus containing between about 1 mg to about 100 mg ofthe active ingredient.

The desired dose may conveniently be presented in a single dose or asdivided doses administered at appropriate intervals, for example, astwo, three, four, or more sub-doses per day. The sub-dose itself may befurther divided, e.g., into a number of discrete loosely spacedadministrations; such as multiple inhalations from an insufflator or byapplication of a plurality of drops into the eye.

The disclosed method can include a kit comprising a compound of theinvention and instructional material which can describe administeringthe compound or a composition comprising the compound to a cell or asubject. This should be construed to include other embodiments of kitsthat are known to those skilled in the art, such as a kit comprising a(preferably sterile) solvent for dissolving or suspending the compoundor composition prior to administering the compound or composition to acell or a subject. Preferably, the subject can be a human.

In accordance with the disclosed methods, as described above or asdiscussed in the Examples below, there can be employed conventionalchemical, cellular, histochemical, biochemical, molecular biology,microbiology, and in vivo techniques which are known to those of skillin the art. Such techniques are explained fully in the literature.

EXAMPLES Example 1:8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid Step 1: 5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-amine

Into a solution of 6-aminonaphthalen-1-ol (1.59 g, 10 mmol, 1.0 eq) inDMF (20 mL) were added cis-4-(tert-butyl)cyclohexyl methanesulfonate(3.04 g, 13 mmol, 1.3 eq) and NaOH (0.60 g, 15 mmol, 1.5 eq). Themixture was stirred at 80° C. for 16 h. After cooling down to rt, themixture was diluted with brine (50 mL) and extracted with EtOAc (60mL×3). The combined organic phase was dried over Na₂SO₄ and concentratedunder reduced pressure. The residue was purified by silica gel plate(EtOAc/petroleum ether=1:10) to give5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-amine as brown oil(1.48 g, Y: 50%).

¹H NMR (400 MHz, CDCl₃) δ: 8.08 (d, J=8.8 Hz, 1H), 7.23 (d, J=8.0 Hz,1H), 7.15 (d, J=8.4 Hz, 1H), 6.95 (s, 1H), 6.90 (dd, J=2.0 Hz, 8.8 Hz,1H), 6.63 (d, J=7.6 Hz, 1H), 4.32-4.23 (m, 1H), 3.00 (br, 2H), 2.31-2.78(m, 2H), 2.16-2.11 (m, 2H), 1.89-1.84 (m, 2H), 1.65-1.11 (m, 3H), 0.89(s, 9H); ESI-MS (M+H)⁺: 298.1.

Step 2: 1-((trans-4-(tert-butyl)cyclohexyl)oxy)-6-iodonaphthalene

Into a solution of5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-amine (900 mg, 3.03mmol, 1.0 eq) in acetonitrile (50 mL) was added 1N HCl (9 mL, 9 mmol,3.0 eq) at 0° C. Then a solution of NaNO₂ (228 mg, 3.3 mmol, 1.1 eq) in10 mL water was added. After stirring at 0° C. for 0.5 h, KI (1.5 g, 9.0mmol, 3.0 eq) was added. The mixture was stirred at rt for another 3 hand diluted with brine (50 mL). The mixture was extracted with EtOAc (80mL×2). The combined organic phase was dried over Na₂SO₄ and concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (petroleum ether) to give1-((trans-4-(tert-butyl)cyclohexyl)oxy)-6-iodonaphthalene as brown oil(310 mg, Y: 25%).

¹H NMR (400 MHz, CDCl₃) δ: 8.05 (s, 1H), 7.87 (d, J=9.2 Hz, 1H), 7.55(dd, J=1.6 Hz, 8.8 Hz, 1H), 7.21 (d, J=8.0 Hz, 1H), 7.12 (d, J=8.0 Hz,1H), 6.73 (d, J=8.0 Hz, 1H), 4.21-4.13 (m, 1H), 2.19-2.16 (m, 2H),1.78-1.75 (m, 2H), 1.43-1.33 (m, 2H), 1.05-0.98 (m, 3H), 0.80 (s, 9H);ESI-MS (M+H)⁺: 409.1.

Step 3: 5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde

Into a solution of1-((trans-4-(tert-butyl)cyclohexyl)oxy)-6-iodonaphthalene (200 mg, 0.49mmol) in dry THF (5 mL) at −78° C. was added n-BuLi (0.38 mL, 2 M inhexane, 1.5 eq). After stirring for 5 min, dry DMF (180 mg, 2.5 mmol,5.0 eq) was added via a syringe. The reaction was allowed to warm to−20° C. and stirred for 30 min. The reaction was quenched with sat. aq.NH₄Cl (10 mL) and extracted with EtOAc (10 mL×2). The combined organicswere dried and concentrated. The residue was purified by columnchromatography on silica gel (petroleum ether) to give compound5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde (80 mg Y: 49%)as a white solid.

¹H NMR (400 MHz, CDCl₃) δ: 10.16 (s, 1H), 8.37 (d, J=8.8 Hz, 1H), 8.28(s, J=1.6 Hz, 1H), 7.90 (dd, J=1.6 Hz, 8.8 Hz, 1H), 7.55 (d, J=8.0 Hz,1H), 7.48-7.45 (m, 1H), 7.02 (d, J=7.6 Hz, 1H), 4.37-4.31 (m, 1H),2.33-2.30 (m, 2H), 1.93-1.90 (m, 2H), 1.59-1.54 (m, 2H), 1.28-1.13 (m,3H), 0.90 (s, 9H); ESI-MS (M+H)⁺: 311.2.

Step 4: methyl8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylate

A mixture of 5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde(93 mg, 0.3 mmol), methyl 8-aza-bicyclo[3.2.1]octane-3-carboxylatehydrochloride (75 mg, 0.24 mmol, 0.8 eq), TEA (30 mg, 0.3 mmol, 1 eq),Na₂SO₄ (210 mg, 1.5 mmol, 5 eq) and NaBH(OAc)₃ (127 mg, 0.6 mmol, 2 eq)in DCM (5 mL) was stirred at rt for 16 h. The mixture was partitionedbetween water (50 mL) and EtOAc (50 mL). The organic layer was washedwith brine (100 mL), dried over Na₂SO₄ and concentrated to give methyl8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylateas a white solid (100 mg, Y: 72%).

¹H NMR (400 MHz, CDCl₃) δ: 8.33 (d, J=8.4 Hz, 1H), 7.84 (s, 1H), 7.53(d, J=8.4 Hz, 1H), 7.44-7.38 (m, 2H), 6.92 (d, J=6.4 Hz, 1H), 4.34-4.29(m, 3H), 3.94-3.93 (m, 2H), 3.67 (s, 3H), 2.77-2.70 (m, 1H), 2.48-2.28(m, 6H), 2.00-1.86 (m, 6H), 1.57-1.48 (m, 2H), 1.22-1.12 (m, 3H), 0.90(s, 9H); ESI-MS (M+H)⁺: 464.3

Step 5:8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid

Into a solution of methyl8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylate(100 mg, 0.22 mmol, 1.0 eq) in MeOH (10 mL) was added NaOH (17 mg, 0.43mmol, 2.0 eq). The mixture was stirred at reflux for 1 h. After coolingdown to rt, the mixture was concentrated and the residue was acidifiedto pH=6 with 1 N HCl. The solid was collected by filtration and washedwith water. After dried under vacuum,8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid was obtained as a white solid (80 mg, Y: 82%).

¹H NMR (400 MHz, CDCl₃) δ: 8.34 (d, J=9.2 Hz, 1H), 7.99 (s, 1H), 7.56(dd, J=1.6 Hz, 8.8 Hz, 1H), 7.49-7.46 (m, 2H), 7.06 (t, J=4.4 Hz, 1H),4.46-4.36 (m, 3H), 4.03-4.01 (m, 2H), 3.00-2.96 (m, 1H), 2.52-2.50 (m,2H), 2.34-2.32 (m, 2H), 2.16-2.10 (m, 5H), 1.96-1.93 (m, 2H), 1.58-1.49(m, 2H), 1.33-1.14 (m, 4H), 0.93 (s, 9H); ESI-MS (M+H)⁺: 450.0, HPLC:100.00%-100.00%.

Example 2:9-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylicacid Step 1: methyl9-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylate

Methyl9-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylatewas obtained following the same procedure as synthesis of methyl8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylateas yellow solid, 80 mg, Y: 56%. ESI-MS (M+H)⁺: 478.3.

Step 2:9-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylicacid

9-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylicacid was obtained following the same procedure as synthesis of8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid as yellow solid, 55 mg, Y: 77%.

¹H NMR (400 MHz, CDCl₃) δ: 8.32 (d, J=8.4 Hz, 1H), 8.04 (s, 1H), 7.61(d, J=8.8 Hz, 1H), 7.47-7.46 (m, 2H), 7.04 (t, J=4.4 Hz, 1H), 4.66 (s,2H), 4.45-4.40 (m, 1H), 3.57-3.56 (m, 2H), 3.14-3.07 (m, 1H), 2.40-2.31(m, 6H), 2.15-2.08 (m, 3H), 1.96-1.75 (m, 5H), 1.58-1.48 (m, 2H),1.33-1.14 (m, 3H), 0.93 (s, 9H); ESI-MS (M+H)⁺: 464.0.

Example 3:3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid Step 1: 5-hydroxy-2-naphthaldehyde

5-hydroxy-2-naphthaldehyde was obtained following the same procedure assynthesis of 5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde asyellow solid, 600 mg, Y: 30%.

¹H NMR (400 MHz, CDCl₃) δ: 10.17 (s, 1H), 8.33-8.30 (m, 2H), 7.95 (d,J=8.8 Hz, 1H), 7.60 (d, J=8.0 Hz, 1H), 7.42 (t, J=8.0 Hz, 1H), 6.99 (d,J=7.6 Hz, 1H), 5.50 (br s, 1H); ESI-MS (M+H)⁺: 173.1.

Step 2: 5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde

A mixture of 5-hydroxy-2-naphthaldehyde (340 mg, 2.0 mmol, 1.0 eq),cis-4-tert-butylcyclohexyl methanesulfonate (700 mg, 3.0 mmol, 1.5 eq)and Cs₂CO₃ (1.30 g, 4.0 mmol, 2.0 eq) in t-BuOH (5 mL) was heated at 80°C. for 16 h and cooled to rt. The mixture was partitioned between EtOAc(20 mL) and water (20 mL). The organic was dried and concentrated. Theresidue was purified by pre-TLC (petroleum ether/EtOAc=20:1) to give5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde (300 mg, Y:50%) as a yellow oil.

¹H NMR (400 MHz, CDCl₃) δ: 10.16 (s, 1H), 8.38 (d, J=8.8 Hz, 1H), 8.28(d, J=1.6 Hz, 1H), 7.90 (dd, J=1.6 Hz, 8.8 Hz, 1H), 7.55 (d, J=8.0 Hz,1H), 7.47 (t, J=8.0 Hz, 1H), 7.02 (d, J=8.0 Hz, 1H), 4.38-4.30 (m, 1H),2.33-2.30 (m, 2H), 1.93-1.90 (m, 2H), 1.58-1.51 (m, 2H), 1.21-1.13 (m,3H), 0.90 (s, 9H); ESI-MS (M+Na)⁺: 333.2.

Step 3: ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylate

A mixture of 5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde(62 mg, 0.2 mmol, 1.0 eq), ethyl3-amino-2,2-dimethylcyclobutanecarboxylate (50 mg, 0.3 mmol, 1.5 eq),HOAc (36 mg, 0.6 mmol, 3.0 eq) and NaBH(OAc)₃ (85 mg, 0.4 mmol, 2.0 eq)in DCM (3 mL) was heated at reflux for 16 h. The mixture was partitionedbetween water (20 mL) and EtOAc (20 mL). The organic layer was washedwith brine (20 mL) and concentrated to give a crude product, which waspurified by pre-TLC (petroleum ether/EtOAc=2:1) to give ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylateas a yellow oil (30 mg, Y: 30%). ESI-MS (M+H)⁺: 466.4.

Step 4:3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid

Into a solution of ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylate(30 mg, 0.08 mmol, 1.0 eq) in MeOH/H₂O (4:1, 2 mL) was added NaOH (16mg, 0.40 mmol, 5.0 eq). The mixture was heated at reflux for 2 h. Aftercooling to rt, the mixture was concentrated and the residue was adjustedto pH=6 with 1 N HCl. The solid was purified by pre-HPLC (ACN and H₂O asmobile phase) to give3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (8 mg, Y: 30%).

¹H NMR (400 MHz, CD₃OD) δ: 8.16 (d, J=8.4 Hz, 1H), 7.78 (s, 1H), 7.40(dd, J=1.6 Hz, 8.4 Hz, 1H), 7.33-7.31 (m, 2H), 6.90-6.87 (m, 1H),4.31-4.25 (m, 1H), 4.09 (AB, 2H), 3.17 (t, J=6.8 Hz, 1H), 2.44 (t, J=7.6Hz, 1H), 2.28-2.19 (m, 3H), 2.12-2.05 (m, 1H), 1.82-1.80 (m, 2H),1.45-1.36 (m, 2H), 1.23 (s, 3H), 1.20-1.10 (m, 3H), 1.05 (s, 3H), 0.81(s, 9H); ESI-MS (M+H)⁺: 438.3.

Example 4:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid Step 1: methyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylatewas obtained following the same procedure as ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylateas a yellow oil (10 mg, Y: 10%). ESI-MS (M+H)⁺: 410.3.

Step 2:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid

1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow oil (4 mg, Y: 20%).

¹H NMR (400 MHz, CD₃OD) δ: 8.16 (d, J=8.8 Hz, 1H), 7.77 (s, 1H),7.34-7.32 (m, 3H), 6.93-6.88 (m, 1H), 4.34-4.27 (m, 3H), 4.00-3.98 (m,4H), 3.29-3.25 (m, 1H), 2.22-2.20 (m, 2H), 1.83-1.80 (m, 2H), 1.45-1.36(m, 2H), 1.20-1.05 (m, 3H), 0.82 (s, 9H); ESI-MS (M+H)⁺: 396.1.

Example 5:2-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)octahydrocyclopenta[c]pyrrole-5-carboxylicacid Step 1: methyl2-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)octahydrocyclopenta[c]pyrrole-5-carboxylate

methyl2-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)octahydrocyclopenta[c]pyrrole-5-carboxylatewas obtained following the same procedure as ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylateas a yellow oil (30 mg, Y: 25%). ESI-MS (M+H)⁺: 464.4.

Step 2:-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)octahydrocyclopenta[c]pyrrole-5-carboxylicacid

2-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)octahydrocyclopenta[c]pyrrole-5-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow solid (10 mg, Y: 30%).

¹H NMR (400 MHz, CD₃OD) δ: 8.10 (d, J=8.8 Hz, 1H), 7.69 (s, 1H), 7.39(dd, J=1.6 Hz, 8.8 Hz, 1H), 7.31-7.25 (m, 2H), 6.84 (dd, J=1.6 Hz, 6.4Hz, 1H), 4.30-4.23 (m, 1H), 3.92 (s, 2H), 2.78-2.54 (m, 7H), 2.22-2.18(m, 2H), 2.07-2.00 (m, 2H), 1.82-1.78 (m, 2H), 1.65-1.59 (m, 2H),1.45-1.35 (m, 2H), 1.19-1.00 (m, 3H), 0.81 (s, 9H); ESI-MS (M+H)⁺:450.3.

Example 6:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)pyrrolidine-3-carboxylicacid Step 1: methyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)pyrrolidine-3-carboxylate

Methyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)pyrrolidine-3-carboxylatewas obtained following the same procedure as ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylateas a yellow oil (20 mg, Y: 25%). ESI-MS (M+H)⁺: 424.4.

Step 2:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)pyrrolidine-3-carboxylicacid

1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)pyrrolidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow solid (6 mg, Y: 30%).

¹H NMR (400 MHz, CD₃OD) δ: 8.15 (d, J=8.4 Hz, 1H), 7.78 (s, 1H), 7.39(dd, J=1.6 Hz, 8.4 Hz, 1H), 7.32-7.31 (m, 2H), 6.89-6.87 (m, 1H),4.31-4.15 (m, 3H), 3.22-3.20 (m, 2H), 3.10-3.06 (m, 2H), 2.99-2.91 (m,1H), 2.22-2.10 (m, 4H), 1.83-1.80 (m, 2H), 1.45-1.36 (m, 2H), 1.20-1.01(m, 3H), 0.81 (s, 9H); ESI-MS (M+H)⁺: 410.3.

Example 7:3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)cyclobutanecarboxylicacid Step 1: ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)cyclobutanecarboxylate

Ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)cyclobutanecarboxylatewas obtained following the same procedure as ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylateas a yellow oil (30 mg, Y: 35%). ESI-MS (M+H)⁺: 438.4.

Step 2:3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)cyclobutanecarboxylicacid

3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)cyclobutanecarboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow solid (2 mg, Y: 10%).

¹H NMR (400 MHz, CD₃OD) δ: 8.16 (d, J=8.8 Hz, 1H), 7.76 (s, 1H), 7.38(dd, J=1.6 Hz, 8.8 Hz, 1H), 7.33-7.31 (m, 2H), 6.91-6.86 (m, 1H),4.34-4.26 (m, 1H), 4.05 (s, 2H), 3.51-3.47 (m, 1H), 2.74-2.67 (m, 1H),2.49-2.42 (m, 2H), 2.23-2.09 (m, 4H), 1.84-1.82 (m, 2H), 1.46-1.37 (m,2H), 1.18-1.02 (m, 3H), 0.82 (s, 9H); ESI-MS (M+H)⁺: 410.3.

Example 8:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azepane-4-carboxylicacid Step 1: methyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azepane-4-carboxylate

Methyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azepane-4-carboxylatewas obtained following the same procedure as ethyl3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylateas a yellow oil (30 mg, Y: 30%). ESI-MS (M+H)⁺: 452.4.

Step 2:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azepane-4-carboxylicacid

1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azepane-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow solid (5 mg, Y: 15%).

¹H NMR (400 MHz, CD₃OD) δ: 8.13 (d, J=8.8 Hz, 1H), 7.74 (s, 1H), 7.41(d, J=8.4 Hz, 1H), 7.32-7.30 (m, 2H), 6.87 (d, J=5.6 Hz, 1H), 4.32-4.26(m, 1H), 4.08 (AB, 2H), 3.16-2.86 (m, 4H), 2.58-2.55 (m, 1H), 2.22-2.19(m, 2H), 2.01-1.68 (m, 8H), 1.46-1.37 (m, 2H), 1.20-0.97 (m, 3H), 0.82(s, 9H); ESI-MS (M+H)⁺: 438.2.

Example 9:1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-hydroxynaphthalen-2-yl)methyl)piperidine-4-carboxylate

A mixture of 5-hydroxy-2-naphthaldehyde (500 mg, 3.0 mmol, 1.0 eq),ethyl piperidine-4-carboxylate (700 mg, 4.5 mmol, 1.5 eq), HOAc (540 mg,9.0 mmol, 3.0 eq) and NaBH(OAc)₃ (1.2 g, 6.0 mmol, 2.0 eq) in DCM (15mL) was stirred at rt for 16 h. The mixture was partitioned betweenwater (200 mL) and EtOAc (200 mL). The organic layer was washed withbrine (100 mL), dried over Na₂SO₄ and concentrated to yield a crudeproduct, which was used for next step without further purification (420mg, Y: 60%). ESI-MS (M+H)⁺: 314.2.

Step 2: ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

A mixture of ethyl1-((5-hydroxynaphthalen-2-yl)methyl)piperidine-4-carboxylate (93 mg, 0.3mmol, 1.0 eq), cis-4-(trifluoromethyl)cyclohexyl methanesulfonate (120mg, 0.5 mmol, 1.5 eq) and Cs₂CO₃ (200 mg, 0.6 mmol, 2.0 eq) in t-BuOH (2mL) was heated at 80° C. for 16 h and cooled to rt. The mixture wasfiltered and the filtrate was purified by pre-TLC (petroleumether/EtOAc=5:1) to give ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate(50 mg, Y: 50%) as a yellow oil. ESI-MS (M+H)⁺: 464.3.

Step 3:1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (20 mg, Y: 50%).

¹H NMR (400 MHz, CD₃OD) δ: 8.12 (d, J=8.8 Hz, 1H), 7.74 (s, 1H), 7.38(d, J=8.4 Hz, 1H), 7.34-7.29 (m, 2H), 6.90 (dd, J=2.0 Hz, 6.4 Hz, 1H),4.38-4.32 (m, 1H), 3.97 (s, 2H), 3.10-3.07 (m, 2H), 2.56-2.53 (m, 2H),2.23-2.10 (m, 4H), 1.94-1.74 (m, 6H), 1.52-1.37 (m, 4H); ESI-MS (M+H)⁺:436.2.

Example 10:1-((5-((cis-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-((cis-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

ethyl1-((5-((cis-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (40 mg, Y: 40%). ESI-MS (M+H)⁺: 464.3.

Step 2:1-((5-((cis-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-((cis-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (20 mg, Y: 50%).

¹H NMR (400 MHz, CD₃OD) δ: 8.22 (d, J=8.8 Hz, 1H), 7.82 (s, 1H), 7.44(dd, J=1.2 Hz, 8.4 Hz, 1H), 7.36-7.34 (m, 2H), 6.92-6.89 (m, 1H),4.80-4.78 (m, 1H), 4.17 (s, 2H), 3.26-3.20 (m, 2H), 2.82-2.77 (m, 2H),2.26-2.14 (m, 4H), 1.96-1.92 (m, 2H), 1.83-1.62 (m, 8H); ESI-MS (M+H)⁺:436.2.

Example 11:1-((5-((cis-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-((cis-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

ethyl1-((5-((cis-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (30 mg, Y: 25%). ESI-MS (M+H)⁺: 438.4.

Step 2:1-((5-((cis-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-((cis-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (13 mg, Y: 30%).

¹H NMR (400 MHz, CD₃OD) δ: 8.23 (d, J=8.8 Hz, 1H), 7.81 (s, 1H), 7.42(dd, J=1.6 Hz, 8.4 Hz, 1H), 7.33-7.30 (m, 2H), 6.86-6.84 (m, 1H), 4.70(s, 1H), 4.16 (s, 2H), 3.24-3.21 (m, 2H), 2.81-2.76 (m, 2H), 2.39-2.35(m, 1H), 2.17-1.88 (m, 6H), 1.67-1.45 (m, 7H), 1.24-1.19 (m, 1H), 0.91(d, J=6.8 Hz, 6H); ESI-MS (M+H)⁺: 410.3.

Example 12:1-((5-((cis-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-((cis-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

Ethyl1-((5-((cis-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (20 mg, Y: 20%). ESI-MS (M+H)⁺: 424.3.

Step 2:1-((5-((cis-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-((cis-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (6 mg, Y: 30%).

¹H NMR (400 MHz, CD₃OD) δ: 8.19 (d, J=8.4 Hz, 1H), 7.72 (s, 1H), 7.39(dd, J=1.6 Hz, 8.4 Hz, 1H), 7.33-7.27 (m, 2H), 6.86-6.81 (m, 1H), 4.72(s, 1H), 3.91 (s, 2H), 3.08-3.05 (m, 2H), 2.48-4.42 (m, 2H), 2.18-2.03(m, 4H), 1.89-1.69 (m, 4H), 1.62-1.51 (m, 4H), 1.43-1.34 (m, 2H),1.25-1.22 (m, 2H), 0.84 (t, J=7.2 Hz, 3H); ESI-MS (M+H)⁺: 396.3.

Example 13:1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

Ethyl1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (20 mg, Y: 20%). ESI-MS (M+H)⁺: 438.3.

Step 2:1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (7 mg, Y: 35%).

¹H NMR (400 MHz, CD₃OD) δ: 8.16 (d, J=8.4 Hz, 1H), 7.78 (s, 1H), 7.38(dd, J=1.2 Hz, 8.8 Hz, 1H), 7.33-7.30 (m, 2H), 6.92-6.87 (m, 1H),4.33-4.26 (m, 1H), 4.14 (s, 2H), 3.23-3.20 (m, 2H), 2.80-2.75 (m, 2H),2.28-2.16 (m, 3H), 1.95-1.74 (m, 6H), 1.45-1.36 (m, 3H), 1.18-1.04 (m,3H), 0.82 (d, J=6.8 Hz, 6H); ESI-MS (M+H)⁺: 410.3.

Example 14:1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

Ethyl1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (20 mg, Y: 10%). ESI-MS (M+H)⁺: 424.3.

Step 2:1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacias a white solid (10 mg, Y: 50%).

¹H NMR (400 MHz, CD₃OD) δ: 8.13 (d, J=8.4 Hz, 1H), 7.73 (s, 1H), 7.36(dd, J=1.6 Hz, 8.8 Hz, 1H), 7.31-7.28 (m, 2H), 6.89-6.86 (m, 1H),4.35-4.27 (m, 1H), 3.97 (s, 2H), 3.11-3.09 (m, 2H), 2.58-2.52 (m, 2H),2.16-2.13 (m, 3H), 1.89-1.71 (m, 6H), 1.48-1.38 (m, 2H), 1.21-0.96 (m,5H), 0.83 (t, J=7.2 Hz, 3H); ESI-MS (M+H)⁺: 396.3.

Example 15:1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

Ethyl1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (20 mg, Y: 10%). ESI-MS (M+H)⁺: 410.3.

Step 2:1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (6 mg, Y: 30%).

¹H NMR (400 MHz, CDCl₃) δ: 8.23 (d, J=8.4 Hz, 1H), 7.73 (s, 1H), 7.45(d, J=8.8 Hz, 1H), 7.36-7.34 (m, 2H), 6.86-6.84 (m, 1H), 4.35-4.30 (m,1H), 4.03 (s, 2H), 3.24-3.20 (m, 2H), 2.40-2.20 (m, 5H), 2.04-1.80 (m,6H), 1.60-1.44 (m, 3H), 1.14-1.03 (m, 2H), 0.93 (d, J=6.4 Hz, 3H);ESI-MS (M+H)⁺: 382.2.

Example 16:1-((5-((4,4-dimethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-((4,4-dimethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

Ethyl1-((5-((4,4-dimethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (30 mg, Y: 25%). ESI-MS (M+H)⁺: 424.3.

Step 2:1-((5-((4,4-dimethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-((4,4-dimethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow oil (10 mg, Y: 35%).

¹H NMR (400 MHz, CD₃OD) δ: 8.20 (d, J=9.2 Hz, 1H), 7.80 (s, 1H), 7.40(dd, J=1.6 Hz, 8.8 Hz, 1H), 7.36-7.30 (m, 2H), 6.91-6.86 (m, 1H),4.49-4.45 (m, 1H), 4.16 (s, 2H), 3.24-3.22 (m, 2H), 2.82-2.76 (m, 2H),2.28-2.23 (m, 1H), 1.96-1.69 (m, 8H), 1.53-1.47 (m, 2H), 1.28-1.21 (m,2H), 0.90 (d, J=7.6 Hz, 6H); ESI-MS (M+H)⁺: 396.3.

Example 17:1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid Step 1: methyl1-((5-hydroxynaphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl 1-((5-hydroxynaphthalen-2-yl)methyl)azetidine-3-carboxylate wasobtained following the same procedure as ethyl1-((5-hydroxynaphthalen-2-yl)methyl)piperidine-4-carboxylate as a yellowsolid (350 mg, Y: 55%). ESI-MS (M+H)⁺: 272.1.

Step 2: methyl1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (15 mg, Y: 11%). ESI-MS (M+H)⁺: 368.3.

Step 3:1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid

1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow oil (10 mg, Y: 75%).

¹H NMR (400 MHz, CD₃OD) δ: 8.25 (d, J=8.8 Hz, 1H), 7.85 (s, 1H),7.46-7.42 (m, 3H), 7.01-6.99 (m, 1H), 4.47-4.40 (m, 1H), 4.29 (s, 2H),4.01-4.00 (m, 4H), 3.38-3.34 (m, 1H), 2.25-2.23 (m, 2H), 1.86-1.83 (m,2H), 1.62-1.47 (m, 3H), 1.23-1.13 (m, 2H), 0.97 (d, J=6.4 Hz, 3H);ESI-MS (M+H)⁺: 354.2.

Example 18:1-((5-((trans-4-phenylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid Step 1: methyl1-((5-((trans-4-phenylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl1-((5-((trans-4-phenylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a white solid (15 mg, Y: 11%). ESI-MS (M+H)⁺: 430.2.

Step 2:1-((5-((trans-4-phenylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid

1-((5-((trans-4-phenylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (10 mg, Y: 69%).

¹H NMR (400 MHz, CD₃OD) δ: 8.30 (d, J=9.2 Hz, 1H), 7.88 (s, 1H),7.48-7.45 (m, 3H), 7.31-7.27 (m, 4H), 7.20-7.17 (m, 1H), 7.10-7.07 (m,1H), 4.61-4.59 (m, 1H), 4.36 (s, 2H), 4.09-4.04 (m, 4H), 3.43-3.36 (m,1H), 2.70-2.63 (m, 1H), 2.41-2.40 (m, 2H), 2.03-2.00 (m, 2H), 1.78-1.69(m, 4H); ESI-MS (M+H)⁺: 416.2.

Example 19:1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid Step 1: methyl1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a white solid (50 mg, Y: 25%). ESI-MS (M+H)⁺: 382.1.

Step 2:1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid

1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (10 mg, Y: 20%).

¹H NMR (400 MHz, CD₃OD) δ: 8.26 (d, J=8.8 Hz, 1H), 7.85 (s, 1H),7.45-7.40 (m, 3H), 7.02-6.97 (m, 1H), 4.47-4.40 (m, 1H), 4.31 (s, 2H),4.07-3.98 (m, 4H), 3.41-3.35 (m, 1H), 2.28-2.25 (m, 2H), 1.93-1.90 (m,2H), 1.60-1.50 (m, 2H), 1.35-1.09 (m, 5H), 0.95 (t, J=7.2 Hz, 3H);ESI-MS (M+H)⁺: 368.3.

Example 20:1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid Step 1: methyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a white solid (15 mg, Y: 11%). ESI-MS (M+H)⁺: 422.2.

Step 2:1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid

1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (5 mg, Y: 34%).

¹H NMR (400 MHz, CD₃OD) δ: 8.16 (d, J=8.8 Hz, 1H), 7.72 (s, 1H),7.41-7.35 (m, 3H), 6.97 (dd, J=1.6 Hz, 8.8 Hz, 1H), 4.48-4.43 (m, 1H),3.89 (s, 2H), 3.66 (t, J=8.0 Hz, 2H), 3.55 (t, J=8.0 Hz, 2H), 3.26-3.22(m, 1H), 2.38-2.23 (m, 3H), 2.07-2.04 (m, 2H), 1.65-1.50 (m, 4H); ESI-MS(M+H)⁺: 408.1.

Example 21:1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid Step 1: methyl1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a white solid (13 mg, Y: 11%). ESI-MS (M+H)⁺: 396.3.

Step 2:1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid

1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a white solid (9 mg, Y: 72%).

¹H NMR (400 MHz, CD₃OD) δ: 8.14 (d, J=8.8 Hz, 1H), 7.68 (s, 1H),7.37-7.34 (m, 3H), 6.90 (dd, J=2.0 Hz, 6.4 Hz, 1H), 4.40-4.34 (m, 1H),3.76 (s, 2H), 3.55 (t, J=8.0 Hz, 2H), 3.38 (t, J=8.0 Hz, 2H), 3.26-3.21(m, 1H), 2.29-2.27 (m, 2H), 1.89-1.85 (m, 2H), 1.56-1.47 (m, 3H),128-1.15 (m, 3H), 0.92 (d, J=6.8 Hz, 6H); ESI-MS (M+H)⁺: 382.3.

Example 22:1-((5-((trans-4-butylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid Step 1: methyl1-((5-((trans-4-butylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl1-((5-((trans-4-butylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (30 mg, Y: 25%). ESI-MS (M+H)⁺: 410.2.

Step 2:1-((5-((trans-4-butylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid

1-((5-((trans-4-butylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow oil (5 mg, Y: 25%).

¹H NMR (400 MHz, CD₃OD) δ: 8.04 (d, J=9.2 Hz, 1H), 7.58 (s, 1H),7.28-7.22 (m, 3H), 6.80 (dd, J=2.0 Hz, 6.8 Hz, 1H), 4.34-4.26 (m, 1H),3.66 (s, 2H), 3.47-3.43 (m, 2H), 3.30-3.25 (m, 4H), 3.16-3.07 (m, 1H),2.18-2.14 (m, 2H), 1.81-1.78 (m, 2H), 1.49-1.39 (m, 2H), 1.27-1.22 (m,5H), 1.08-0.98 (m, 2H), 0.83 (t, J=7.2 Hz, 3H); ESI-MS (M+H)⁺: 396.3.

Example 23:1-((5-((trans-4-cyclopentylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid Step 1: methyl1-((5-((trans-4-cyclopentylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylate

Methyl1-((5-((trans-4-cyclopentylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylatewas obtained following the same procedure as ethyl1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a yellow oil (25 mg, Y: 20%). ESI-MS (M+H)⁺: 422.1.

Step 2:1-((5-((trans-4-cyclopentylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid

1-((5-((trans-4-cyclopentylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid was obtained following the same procedure as3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid as a yellow oil (10 mg, Y: 50%).

¹H NMR (400 MHz, CD₃OD) δ: 8.04 (d, J=8.4 Hz, 1H), 7.57 (s, 1H),7.27-7.22 (m, 3H), 6.80 (dd, J=1.6 Hz, 6.4 Hz, 1H), 4.32-4.25 (m, 1H),3.66 (s, 2H), 3.47-3.43 (m, 2H), 3.28-3.26 (m, 2H), 3.15-3.07 (m, 1H),2.16-2.13 (m, 2H), 1.85-1.83 (m, 2H), 1.74-1.67 (m, 2H), 1.55-1.37 (m,8H), 1.09-1.04 (m, 4H); ESI-MS (M+H)⁺: 408.3.

Example 24:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid Step 1: ethyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

A mixture of 5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde(145 mg, 0.47 mmol), ethyl piperidine-4-carboxylate (147 mg, 0.94 mmol,2 eq) and NaBH(OAc)₃ (198 mg, 0.94 mol, 2 eq) in DCE (10 mL) was stirredat rt for 2 h. Water (100 mL) was added to the mixture. The organiclayer was washed with brine (100 mL), dried over Na₂SO₄ and concentratedto give ethyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylateas a white solid (80 mg, Y: 38%). ESI-MS (M+H)⁺: 452.3

¹H NMR (400 MHz, CDCl₃) δ: 8.20 (d, J=8.8 Hz, 1H), 7.66 (s, 1H), 7.44(dd, J=8.8, 1.2 Hz, 1H), 7.37-7.31 (m, 2H), 6.83 (dd, J=6.8, 1.6 Hz,1H), 4.31-4.27 (m, 1H), 4.18 (q, J=7.2 Hz, 2H), 3.64 (s, 2H), 2.90-2.88(m, 2H), 2.33-2.25 (m, 3H), 2.09-2.04 (m, 2H), 1.90-1.87 (m, 5H),1.28-1.08 (m, 9H), 0.89 (s, 9H).

Step 2:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

To a solution of ethyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate(80 mg, 0.18 mmol, 1.0 eq) in MeOH/H₂O (4:1, 10 mL) was added NaOH (14mg, 0.35 mmol, 2.0 eq). The mixture was stirred at reflux for 1 h. Aftercooling down to rt, the mixture was concentrated and the residue wasadjusted to pH=6 with 1 N HCl. The solid was collected by filtration andwashed with water. After dried under vacuum,1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid was obtained as a light yellow solid (34 mg, Y: 45%). ESI-MS(M+H)⁺: 424.3, HPLC: 100.00%-100.00%.

¹H NMR (400 MHz, CDCl₃) δ: 8.20 (d, J=8.0 Hz, 1H), 7.82 (s, 1H), 7.40(dd, J=9.2, 1.6 Hz, 1H), 7.36-7.35 (m, 2H), 6.94 (q, J=4.4 Hz, 1H),4.36-4.29 (m, 1H), 4.23 (s, 2H), 3.30-3.27 (m, 2H), 2.89-2.87 (m, 2H),2.35-2.30 (m, 1H), 2.24-2.22 (m, 2H), 1.99-1.96 (m, 2H), 1.86-1.83 (m,4H), 1.47-1.39 (m, 2H), 1.17-1.04 (m, 3H), 0.90 (s, 9H).

Example 25:1-((5-(heptyloxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic acidStep 1: 6-bromonaphthalen-1-ol

A solution of tetrabutylammonium tribromide (832 mg, 1.1 eq, 1.73 mmol)in DCM (4 mL) was added dropwise to a solution of6-bromo-3,4-dihydronaphthalen-1(2H)-one (350 mg, 1.0 eq, 1.57 mmol) indichloromethane (2 mL) and methanol (2 mL) at room temperature over 1 h.At completion of the addition, the mixture was stirred at rt for 15 hrsand was then concentrated. The residue was taken into DCM and was washedwith saturated sodium bicarbonate three times. The organic layer wasconcentrated and the residue was dissolved in dimethylformamide (10 mL).Lithium carbonate (286 mg, 2.1 eq, 3.29 mmol) and lithium bromide (372mg, 3.2 eq, 5.02 mmol) were added and the resulting mixture was stirredat 140° C. for 1.5 hrs. After cooling to rt, the solids were filteredand rinsed with ethyl acetate. The filtrate was washed with water fourtimes and dried over sodium sulfate to give 6-bromonaphthalen-1-ol as abrown solid (195 mg, Y: 56%).

¹H NMR (400 MHz, CDCl₃) δ: 8.06 (d, J=9.2 Hz, 1H), 7.97 (d, J=2.4 Hz,1H), 7.54 (dd, J=8.8 Hz, 2.0 Hz, 1H), 7.35-7.29 (m, 2H), 6.80 (dd, J=6.0Hz, 1.6 Hz, 1H), 5.20 (s, 1H).

Step 2: 6-bromo-1-(heptyloxy)naphthalene

To a solution of 6-bromonaphthalen-1-ol (156 mg, 0.7 mmol, 1.0 eq) inDMF (5 mL) were added 1-bromoheptane (370 mg, 2.1 mmol, 3.0 eq) andK₂CO₃ (97 mg, 0.7 mmol, 1.0 eq). The mixture was stirred at 80° C. for 2h. After cooling down to rt, the mixture was diluted with brine (50 mL)and extracted with EtOAc (60 mL×3). The combined organic phase was driedover Na₂SO₄ and concentrated under reduced pressure. The residue waspurified by silica gel plate (EA:PE=1:10) to give6-bromo-1-(heptyloxy)naphthalene as brown oil (180 mg, Y: 86%). ESI-MS(M+H)⁺: 321.1.

¹H NMR (400 MHz, CDCl₃) δ: 8.14 (d, J=9.2 Hz, 1H), 7.93 (d, J=2.0 Hz,1H), 7.51 (dd, J=8.8 Hz, 2.0 Hz, 1H), 7.37 (t, J=8.4 Hz, 1H), 7.30-7.28(m, 1H), 6.79 (d, J=7.6 Hz, 1H), 4.11 (t, J=6.4 Hz, 2H), 1.95-1.88 (m,2H), 1.58-1.51 (m, 2H), 1.44-1.34 (m, 6H), 0.92-0.89 (m, 3H).

Step 3: 5-(heptyloxy)-2-naphthaldehyde

5-(heptyloxy)-2-naphthaldehyde was prepared following the same procedureas 5-((trans-4-(tert-butyl)cyclohexyl)oxy)-2-naphthaldehyde, as yellowoil, 130 mg, Y: 83%. ESI-MS (M+H)⁺: 271.2.

¹H NMR (400 MHz, CDCl₃) δ: 10.16 (s, 1H), 8.39 (d, J=8.4 Hz, 1H), 8.28(s, 1H), 7.92 (dd, J=9.2 Hz, 1.6 Hz, 1H), 7.57-7.55 (m, 1H), 7.47 (t,J=7.6 Hz, 1H), 6.95 (d, J=7.6 Hz, 1H), 4.15 (t, J=6.8 Hz, 2H), 1.97-1.90(m, 2H), 1.60-1.53 (m, 2H), 1.45-1.38 (m, 2H), 1.35-1.32 (m, 4H), 0.90(t, J=6.8 Hz, 3H).

Step 4: ethyl1-((5-(heptyloxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate

Ethyl 1-((5-(heptyloxy)naphthalen-2-yl)methyl)piperidine-4-carboxylatewas prepared following the same procedure as ethyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylate,as yellow oil, 170 mg, Y: 86%. ESI-MS (M+H)⁺: 412.3.

¹H NMR (400 MHz, CDCl₃) δ: 8.22 (d, J=8.4 Hz, 1H), 7.68 (s, 1H), 7.46(d, J=8.8 Hz, 1H), 7.37-7.32 (m, 2H), 6.77 (dd, J=6.8 Hz, 2.0 Hz, 1H),4.15-4.09 (m, 4H), 3.65 (s, 2H), 2.91-2.88 (m, 2H), 2.34-2.26 (m, 1H),2.10-2.06 (m, 2H), 1.93-1.80 (m, 6H), 1.59-1.52 (m, 2H), 1.44-1.39 (m,2H), 1.34-1.31 (m, 4H), 1.24 (t, J=7.2 Hz, 3H), 0.90 (t, J=6.8 Hz, 3H).

Step 5: 1-((5-(heptyloxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

1-((5-(heptyloxy)naphthalen-2-yl)methyl)piperidine-4-carboxylic acid wasprepared following the same procedure as1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid, as yellow oil, 55 mg, Y: 74%. ESI-MS (M+H)⁺: 384.3. HPLC:100.00%-100.00%.

¹H NMR (400 MHz, MeOD) δ: 8.30 (d, J=8.8 Hz, 1H), 7.89 (s, 1H), 7.50 (d,J=8.8 Hz, 1H), 7.44-7.43 (m, 2H), 6.92-6.91 (m, 1H), 4.26 (s, 2H), 4.15(t, J=6.4 Hz, 2H), 3.35-3.34 (m, 2H), 2.92-2.85 (m, 2H), 2.40-2.35 (m,1H), 2.06-2.01 (m, 2H), 1.95-1.89 (m, 4H), 1.61-1.54 (m, 2H), 1.46-1.41(m, 2H), 1.36-1.34 (m, 4H), 0.91 (t, J=6.8 Hz, 3H).

Example 26:9-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylicacid Step 1: (5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methanol

To a mixture of 5-(4-methyl-cyclohexyloxy)-naphthalene-2-carbaldehyde(400 mg, 1 mmol) in tetrahydrofuran (7.254 mL, 89.44 mmol) was added1.00M of lithium tetrahydroaluminate in tetrahydrofuran (3.726 mL, 3.726mmol). Gas evolution observed. The reaction was then stirred at rt for30 min, LCMS showed complete conversion. EtOAc was added and rochele'ssalt was added and stirred for 30 min. The organic layer was washed withbrine, dried and evaporated, and dried under high vacuum to give(5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methanol (0.4 g). LCMS:RT 1.92 min; M m/z=271.10 M+1. It was used to next step as is.

Step 2: (5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methylmethanesulfonate

To a solution of [5-(4-methyl-cyclohexyloxy)-naphthalen-2-yl]-methanol(413 mg, 1.53 mmol) and N,N-diisopropylethylamine (0.79822 mL, 4.5827mmol) in methylene chloride (7.1 mL, 110 mmol) was added methanesulfonylchloride (0.23647 mL, 3.0551 mmol) dropwise. A white precipitate formed.The solution was stirred at rt for 5 h. LCMS showed no starting materialleft, and complete conversion to RT 2.39 min m/z=289.00. The mixture wasdiluted with DCM and washed with sodium bicarbonate aq solution andwater, dried over MgSO₄, filtered, concentrated. The residue was used asin the next step.

Step 3:9-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylicacid

To a solution of methanesulfonic acid5-(4-methyl-cyclohexyloxy)-naphthalen-2-ylmethyl ester (177 mg, 0.508mmol) in DMF (1.9665 mL, 25.398 mmol),9-aza-bicyclo[3.3.1]nonane-3-carboxylic acid methyl ester HCl salt(223.20 mg, 1.0159 mmol) was added, followed by cesium carbonate (496.50mg, 1.5238 mmol). The reaction was then heated at 80° C. for 1 h. LCMSshowed no SM left, and the completion of the reaction. After cooleddown, the reaction mixture was diluted with EtOAc, washed with water(2×). The organic phase was then separated, dried and concentrated. Thecrude was purified by HPLC, removed the solvent, the ester was thendissolved in tetrahydrofuran (2.4 mL, 29 mmol), treated with 1.0 M oflithium hydroxide in water (3.5 mL, 3.5 mmol) at rt overnight. Acidifiedwith 1NHCl, the organic layer was dried and concentrated. The crude wasthen purified by HLPC to give9-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylicacid as a white powder (86.9 mg, 41%). LCMS: RT=1.43 min.; MH+ 422.10.

¹H NMR (400 MHz, METHANOL-d4) δ 8.42 (d, J=8.72 Hz, 1H), 8.07 (d, J=4.77Hz, 1H), 7.65 (d, J=8.60 Hz, 1H), 7.49 (d, J=5.46 Hz, 2H), 6.95-7.10 (m,1H), 4.61-4.81 (m, 2H), 3.40-3.79 (m, 2H), 1.35-2.75 (m, 20H), 1.00 (d,J=5.84 Hz, 3H).

Example 27:8-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid

To a solution of methanesulfonic acid5-(4-methyl-cyclohexyloxy)-naphthalen-2-ylmethyl ester (177 mg, 0.508mmol) in DMF (2 mL), 8-aza-bicyclo[3.2.1]octane-3-carboxylic acid methylester HCl salt (208.38 mg, 1.0131 mmol) was added, followed by cesiumcarbonate (495.14 mg, 1.5197 mmol). The reaction was then heated at 80°C. for 1 h. After cooled down, the reaction mixture was diluted withEtOAc, washed with water (2×). The organic phase was then separated,dried and concentrated. The crude was purified by HPLC, removed thesolvent, the ester was then dissolved in tetrahydrofuran (2.3 mL, 29mmol), treated with 1.0 M of lithium hydroxide in Water (3.5 mL, 3.5mmol) at rt overnight. Acidified with 1NHCl, the organic layer was driedand concentrated. The crude was then purified by HLPC to give8-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid as a white powder (132.9 mg, 64%). LCMS: RT 1.40 min.; MH+ 408.1.

¹H NMR (400 MHz, METHANOL-d4) δ 8.38 (d, J=8.72 Hz, 1H), 7.98 (s, 1H),7.60 (d, J=10.48 Hz, 1H), 7.46 (d, J=4.96 Hz, 2H), 6.92-7.08 (m, 1H),4.85 (br. s., 1H), 3.80-4.45 (m, 2H), 3.37 (s, 2H), 1.35-2.81 (m, 18H),1.00 (d, J=5.65 Hz, 3H).

Example 28:1-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid

To a solution of methanesulfonic acid5-(cis-4-methyl-cyclohexyloxy)-naphthalen-2-ylmethyl ester (177 mg,0.508 mmol) in DMF (2 mL), ethyl piperidine-4-carboxylate (159.71 mg,1.0159 mmol) was added, followed by cesium carbonate (496.50 mg, 1.5238mmol).

The reaction was then heated at 80° C. for 1 h. After cooled down, thereaction mixture was diluted with EtOAc, washed with water (2×). Theorganic phase was then separated, dried and concentrated. The crude waspurified by HPLC, removed the solvent, the ester was then dissolved inTHF (2.4 mL, 29 mmol), treated with 1.0 M of lithium hydroxide in water(3.5 mL, 3.5 mmol) at rt overnight. Acidified with 1NHCl, the organiclayer was dried and concentrated. The crude was then purified by HLPC togive1-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid as a white powder (160 mg, 82%). LCMS: RT 1.46 min.; MH+ 382.10.

¹H NMR (400 MHz, METHANOL-d4) δ 8.41 (d, J=8.66 Hz, 1H), 7.98 (s, 1H),7.56 (d, J=10.42 Hz, 1H), 7.49 (s, 2H), 6.92-7.10 (m, 1H), 4.83-4.85 (m,OH), 4.49 (s, 2H), 3.61 (d, J=12.61 Hz, 2H), 2.98-3.19 (m, 2H),1.38-2.75 (m, 14H), 1.00 (d, J=5.84 Hz, 3H).

Example 29:1-((5-(heptyloxy)naphthalen-1-yl)methyl)piperidine-4-carboxylic acidStep 1: of 5-iodonaphthalen-1-ol

To a solution of 5-aminonaphthalen-1-ol (1.59 g, 10 mmol, 1.0 eq) inCH₃CN (100 mL) were added p-TsOH.H₂O (5.7 g, 30 mmol, 3.0 eq) and asolution of NaNO₂ (1.87 g, 27.5 mmol, 1.1 eq) in 10 mL water at 0° C.After stirring at 0° C. for 1 h, KI (25 mmol, 4.15 g, 2.5 eq) was addedand the mixture was stirred at rt for another 3 h. After diluted withEtOAc (200 mL), the mixture was washed with water (80 mL). The organicphase was dried over Na₂SO₄ and concentrated under reduced pressure. Theresidue was purified by silica gel plate (EA:PE=1:10) to give5-iodonaphthalen-1-ol as a brown solid (1.35 g, yield: 50%). ESI-MS(M+H)⁺: 271.1.

¹H NMR (400 MHz, CDCl₃) δ: 8.23 (d, J=8.4 Hz, 1H), 8.09 (dd, J=7.6, 1.2Hz, 1H), 7.69 (d, J=8.4 Hz, 1H), 7.40 (dd, J=8.4, 8.0 Hz, 1H), 7.17 (dd,J=8.4, 7.2 Hz, 1H), 6.87 (d, J=7.2 Hz, 1H).

Step 2: 1-(heptyloxy)-5-iodonaphthalene

To a solution of 5-iodonaphthalen-1-ol (1.35 mg, 5 mmol, 1.0 eq) intoluene (10 mL) were added 1-heptanol (1.74 g, 15 mmol, 3.0 eq), PPh₃(1.97 g, 7.5 mmol, 1.5 eq) and DIAD (1.52 g, 7.5 mmol, 1.5 eq)successively under N₂. The mixture was stirred at rt for 0.5 h and thesolvent was removed under reduced pressure. The residue was purified bysilica gel column (pure petroleum ether) to give1-(heptyloxy)-5-iodonaphthalene as a slight yellow solid (1.1 g, yield:60%). ESI-MS (M+H)⁺: 369.1.

¹H NMR (400 MHz, CDCl₃) δ: 8.24 (d, J=8.4 Hz, 1H), 8.01 (dd, J=7.6, 1.2Hz, 1H), 7.57 (d, J=8.8 Hz, 1H), 7.38 (t, J=8.0 Hz, 1H), 7.07 (dd,J=8.0, 7.2 Hz, 1H), 6.78 (d, J=7.6 Hz, 1H), 4.05 (t, J=6.4 Hz, 2H),1.88-1.81 (m, 2H), 1.51-1.44 (m, 2H), 1.35-1.15 (m, 6H), 0.80 (t, J=7.2Hz, 3H).

Step 3: 5-(heptyloxy)-1-naphthaldehyde

To a solution of 1-(heptyloxy)-5-iodonaphthalene (1.1 mg, 3.0 mmol, 1.0eq) in dry THF (10 mL) was added n-BuLi (2.0 N in THF, 2.25 mL, 4.5mmol, 1.3 eq) at −78° C. under N₂. The mixture was stirred at thistemperature for 0.5 h and then DMF (5.0 mg, 6.9 mmol, 5.0 eq) was added.The resulting mixture was stirred at −78° C. for another 0.5 h. Water(20 mL) was added to quench the reaction and the mixture was extractedwith EtOAc (100 mL×2). The combined organic phases were dried overNa₂SO₄, concentrated and the residue was purified by silica gel column(EA:PE=1:20) to give 5-(heptyloxy)-1-naphthaldehyde as a slight yellowsolid (700 mg, yield: 86%). ESI-MS (M+H)⁺: 311.1.

¹H NMR (400 MHz, CDCl₃) δ: 10.32 (s, 1H), 8.77 (d, J=8.4 Hz, 1H), 8.63(d, J=8.4 Hz, 1H), 8.01 (dd, J=7.2, 1.2 Hz, 1H), 7.63-7.57 (m, 2H), 6.92(d, J=8.0 Hz, 1H), 4.15 (t, J=6.4 Hz, 2H), 1.97-1.92 (m, 2H), 1.59-1.53(m, 2H), 1.44-1.40 (m, 2H), 1.39-1.30 (m, 4H), 0.89 (t, J=6.8 Hz, 3H.

Step 4: ethyl1-((5-(heptyloxy)naphthalen-1-yl)methyl)piperidine-4-carboxylate

The preparation of ethyl1-((5-(heptyloxy)naphthalen-1-yl)methyl)piperidine-4-carboxylate was thesame as that of ethyl1-((6-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylate,130 mg, as yellow oil, yield: 50%. ESI-MS (M+H)⁺: 412.3.

¹H NMR (400 MHz, CDCl₃) δ: 8.27 (d, J=7.6 Hz, 1H), 7.82 (d, J=8.4 Hz,1H), 7.44-7.38 (m, 3H), 6.82 (d, J=8.0 Hz, 1H), 4.07-4.01 (m, 4H), 3.78(s, 2H), 2.86-2.83 (m, 2H), 2.23-2.19 (m, 1H), 2.04-2.00 (m, 2H),1.87-1.68 (m, 6H), 1.52-1.45 (m, 2H), 1.35-1.24 (m, 6H), 1.16 (t, J=7.2Hz, 3H), 0.83 (t, J=6.8 Hz, 3H).

Step 5: 1-((5-(heptyloxy)naphthalen-1-yl)methyl)piperidine-4-carboxylicacid

The preparation of1-((5-(heptyloxy)naphthalen-1-yl)methyl)piperidine-4-carboxylic acid wasthe same as that of1-((6-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylicacid, 13 mg, as white solid, yield: 50%. ESI-MS (M+H)⁺: 384.2, HPLC:100%-100%.

¹H NMR (400 MHz, CD₃OD) δ: 8.45 (d, J=8.4 Hz, 1H), 7.79 (s, 1H), 7.77(d, J=2.0 Hz, 1H), 7.62-7.55 (m, 2H), 7.03 (d, J=8.0 Hz, 1H), 4.80 (s,2H), 4.19 (t, J=6.4 Hz, 2H), 3.53-3.49 (m, 2H), 3.28-3.22 (m, 2H),2.72-2.64 (m, 1H), 2.18-2.15 (m, 2H), 1.99-1.92 (m, 4H), 1.62-1.57 (m,2H), 1.47-1.34 (m, 6H), 0.93 (t, J=6.8 Hz, 3H).

Example 30:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylicacid Step 1: 1-((trans-4-(tert-butyl)cyclohexyl)oxy)-5-iodonaphthalene

The preparation of1-((trans-4-(tert-butyl)cyclohexyl)oxy)-5-iodonaphthalene was the sameas that of 1-(heptyloxy)-5-iodonaphthalene. 1 g, as a slight solid,yield: 50%. ESI-MS (M+H)⁺: 409.1

¹H NMR (400 MHz, CDCl₃) δ: 8.23 (d, J=8.4 Hz, 1H), 7.98 (dd, J=7.2, 0.8Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.37 (t, J=8.0 Hz, 1H), 7.07-7.03 (m,1H), 6.83 (d, J=8.4 Hz, 1H), 4.26-4.21 (m, 1H), 2.24-2.20 (m, 2H),1.83-1.80 (m, 2H), 1.48-1.39 (m, 2H), 1.14-1.03 (m, 3H), 0.80 (s, 9H).

Step 2: 5-((trans-4-(tert-butyl)cyclohexyl)oxy)-1-naphthaldehyde

The preparation of5-((trans-4-(tert-butyl)cyclohexyl)oxy)-1-naphthaldehyde was the same asthat of 5-(heptyloxy)-1-naphthaldehyde. 400 mg, as a yellow solid,yield: 40%. ESI-MS (M+H)⁺: 311.2.

¹H NMR (400 MHz, CDCl₃) δ: 10.40 (s, 1H), 8.75 (d, J=8.8 Hz, 1H), 8.64(d, J=8.4 Hz, 1H), 7.99 (dd, J=6.8, 1.2 Hz, 1H), 7.60-7.55 (m, 2H), 6.97(d, J=8.0 Hz, 1H), 4.38-4.32 (m, 1H), 2.34-2.30 (m, 2H), 1.93-1.89 (m,2H), 1.58-1.49 (m, 2H), 1.25-0.92 (m, 3H), 0.89 (s, 9H).

Step 3: ethyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylate

The preparation of ethyl1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylatewas the same as that of ethyl1-((6-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylate.200 mg, as a white solid, yield: 56%. ESI-MS (M+H)⁺: 452.3.

¹H NMR (400 MHz, CDCl₃) δ: 8.24 (d, J=7.6 Hz, 1H), 7.81 (d, J=8.4 Hz,1H), 7.43-7.36 (m, 3H), 6.87 (d, J=7.6 Hz, 1H), 4.33-4.28 (m, 1H), 4.11(q, J=7.2 Hz, 2H), 3.84 (s, 2H), 2.93-2.89 (m, 2H), 2.33-2.30 (m, 3H),2.12-2.07 (m, 2H), 1.91-1.74 (m, 6H), 1.27-1.13 (m, 5H), 1.25 (t, J=7.2Hz, 3H), 0.86 (s, 9H)

Step 4:1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylicacid

The preparation of1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylicacid was the same as that of1-((6-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylicacid. 70 mg, as a white solid, yield: 50%. ESI-MS (M+H)⁺: 424.3. HPLC:98.54%-100%.

¹H NMR (400 MHz, CD₃OD) δ: 8.37 (d, J=8.4 Hz, 1H), 7.72 (d, J=8.8 Hz,1H), 7.65 (d, J=7.2 Hz, 1H), 7.52-7.44 (m, 2H), 6.98 (d, J=8.0 Hz, 1H),4.52 (s, 2H), 4.39-4.16 (m, 1H), 3.35-3.31 (m, 2H), 2.94-2.85 (m, 2H),2.37-2.30 (m, 3H), 2.02-1.89 (m, 6H), 1.55-1.49 (m, 2H), 1.27-1.13 (m,3H), 0.91 (s, 9H).

Example 31: S1P Receptor Activity Assays

Compounds that are not specific for a particular S1P receptor can causeundesirable side effects. Accordingly, compounds are tested to identifythose that are specific. Accordingly, the test compounds are tested in acalcium mobilization assay/S1P receptor activity assay. The procedure isessentially as described in Davis et al. (2005) Journal of BiologicalChemistry, vol. 280, pp. 9833-9841, which is incorporated by referencein its entirety with the following modifications. Calcium mobilizationassays are performed in recombinant CHEM cells expressing human S1P₁,S1P₂, S1P₃, S1P₄, or S1P₅ purchased from Millipore (Billerica, Mass.).To detect free intracellular calcium, S1P₁, S1P₂, S1P₃, S1P₄, or S1P₅cells are loaded with FLIPR Calcium 4 dye from Molecular Devices(Sunnyvale, Calif.). Cells are imaged for calcium mobilization using aFLIPR^(TETRA) equipped with a 96-well dispense head.

Agonist percentage activation determinations were obtained by assayingsample compounds and referencing the E_(max) control for each receptorprofiled. Antagonist percentage inhibition determinations were obtainedby assaying sample compounds and referencing the control EC₈₀ wells foreach receptor profiled.

Calcium Flux Assay: Agonist Assay Format

Sample compounds were plated in an eight-point, four-fold dilutionseries in duplicate with a top concentration of 10 μM. Theconcentrations described here reflect the final concentration of thecompounds during the antagonist assay. During the agonist assay thecompound concentrations were 1.25 fold higher to allow for the finaldesired concentration to be achieved with further dilution by EC₈₀ ofreference agonists during the antagonist assay.

Reference agonists were handled as mentioned above serving as assaycontrol. The reference agonists were handled as described above forE_(max).

Assay was read for 180 seconds using the FLIPR^(TETRA) (This assay runadded sample compounds and reference agonist to respective wells). Atthe completion of the first “Single Addition” assay run, assay plate wasremoved from the FLIPR^(TETRA) and placed at 25° C. for seven (7)minutes.

Calcium Flux Assay: Antagonist Assay Format

Using the EC₈₀ values determined during the agonist assay, stimulatedall pre-incubated sample compound and reference antagonist (ifapplicable) wells with EC₈₀ of reference agonist. Read for 180 secondsusing the FLIPR^(TETRA) (This assay added reference agonist torespective wells—then fluorescence measurements were collected tocalculate percentage inhibition values).

With regard to S1P4 antagonist activity, the compound of example 30 hadan IC₅₀ value of no greater than 250 nM.

Example 32: ATX Activity Measurements

ATX (Autotaxin) is a 125 KDa glycoprotein with lysophospholipase D(LPLD) activity that generates the bioactive lipid lysophosphatidic acid(LPA) from lysophosphatidylcholine (LPC). The ATX biochemical assayutilizes a FRET (fluorescence resonance energy transfer) technologyplatform. The fluorescence signal of FRET substrate FS-3 is quenched dueto intra-molecular FRET of a fluorophore to a non-fluorescing quencher(Ferguson, C. G., et al., Org Lett. 2006 May 11; 8(10): 2023-2026, whichis incorporated by reference in its entirety). ATX catalyzes thehydrolysis of the substrate which separates the dabsyl quencher from thefluorescein reporter, which becomes fluorescent. The reaction ismonitored by a SpectraMax M5 (Molecular Devices, Sunnyvale, Calif.) withat excitation wavelength 485 nm and emission wavelength 535 nm.

Reagents

Fatty acid free-BSA (Sigma A8806): 10 mg/mL in H₂O, stored at 4° C.

2×ATX assay buffer: 100 mM Tris, 280 mM NaCl, 10 mM KCl, 2 mM CaCl₂, 2mM MgCl₂, pH 7.4.

Human ATX protein: expressed and purified in house. Stored at −80° C.

Substrate FS-3 (Echelon, L-2000): 100 μg in 77.74 μL H₂O (1 mM stock),stored at −20° C.

384-well flat bottom plates—Corning #3575.

Assay

Compound dilution—All compounds were provided at 10 mM in 100% DMSO. Inthe first well, 2 μL of 10 mM compound was added to 78 μL of DMSO (1:40dilution). In subsequent wells 3-fold dilution (total 10 dilutions) wereperformed.

1×ATX assay buffer was made up with a final concentration of 1 mg/mLfatty acid free-BSA using 2×ATX assay buffer, 10 mg/ml fatty acidfree-BSA and ddH₂O.

ATX protein was diluted with 1×ATX assay buffer to a concentration of1.32 μg/mL (1.32×). 38 μL was added per well to the assay plate. Thefinal concentration of ATX in the reaction as 1.0 μg/mL.

2 μL per well of compounds was transferred to provide the desiredconcentration. The plate was centrifuged, then incubated at roomtemperature for 30 minutes on the shaker.

FS-3 was diluted with 1×ATX assay buffer to a concentration of FS-3 of10 μM (5×). Then, 10 μL was added per well to the assay plate. The finalconcentration of FS-3 in the reaction was 2 μM. The plate wascentrifuged. The plate was kept shaking at room temperature for 2 hours.Because FS-3 substrate is light sensitive, plates were kept covered andprotected from light.

Fluorescence was measured using SpectraMax M5 (excitation at 485nm/emission at 538 nm, top read).

With regard to ATX activity, compounds of the invention had thefollowing IC₅₀ values:

IC50 (μM) Example No. Less than 0.5 μM 4, 24 0.5 μM to 5 μM 1, 3, 5, 7,9, 12, 13, 14, 15, 19, 20, 26, 27 Greater than 5 μM 2, 8, 10, 11, 17,18, 21, 22, 23, 25, 28

OPC Differentiation Assay

Enriched populations of oligodendrocytes were grown from post-natal day2 (P2) female Sprague Dawley rats. The forebrain was dissected out andplaced in Hank's buffered saline solution (HBSS; Invitrogen, GrandIsland, N.Y.). The tissue was cut into 1 mm fragments and incubated at37° C. for 15 minutes in 0.01% trypsin and 10 μg/mL DNase. Dissociatedcells were plated on poly-L-lysine-coated T75 tissue culture flasks andgrown at 37° C. for 10 days in Dulbecco's modified Eagle's medium (DMEM)with 20% fetal calf serum (Invitrogen). A2B5⁺ OPCs were collected byshaking the flask overnight at 200 rpm and 37° C., resulting in a 95%pure population.

For the differentiation assay, 2 μM and 20 μM antagonist or the sameconcentrations of vehicle (DMSO) were applied to OPCs cultured inCNTF/T3 containing media. After a 3-day incubation, cells were lysed in80 μL lysis buffer (50 mM HEPES[4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid], pH 7.5, 150 mMNaCl, 1.5 mM MgCl₂, 1 mM ethylene glycol tetraacetic acid [EGTA], 1%Triton X-100 and 10% glycerol) for 30 minutes at 4° C. Aftercentrifugation at 14,000 g for 15 minutes, the supernatants were boiledin Laemmli sample buffer, subjected to 4-20% SDS-PAGE, and analyzed byWestern blotting with anti-MBP, anti-myelin-associated glycoprotein(MAG), or anti-beta actin antibodies. The secondary antibodies used wereanti-mouse IgG-HRP (horseradish peroxidase) and anti-rabbit IgG-HRPrespectively.

DRG-OPC Myelination Assay

Embryonic neocortical neurons are dissected from embryonic day 18 (E18)Sprague Dawley rats, and then plated on poly-D-lysine (100 μg/mL)-coatedcover slips and grown in neurobasal medium supplemented with B27(Invitrogen) for one week. A2B5⁺ OPCs are prepared as described aboveand then added into the cultured neocortical neurons. One day later,different concentrations of an S1P4 receptor antagonist or ATX inhibitorand control reagents are applied into the co-cultures. Fresh mediacontaining the different concentrations of an S1P4 receptor antagonistor ATX inhibitor or control compounds are supplied every three days.After ten days, co-cultures are subjected to sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE)/Western blot analyses toquantify MAG, MBP, and MOG.

Remyelination Assay in Brain Slice Culture

Approximately three to four consecutive 300 μm slices are taken from thejunction of the corpus callosum to the hippocampus in post-natal, day 17Sprague Dawley rats (Charles River, Willmington, Mass.). Slices arecultured in basal DMEM supplemented with 25% horse serum for three days,before being treated with 6 mg/mL LPC (Sigma L-4129) for a further threedays. The medium is then changed, and slices incubated with mediumcontaining an S1P4 receptor antagonist or ATX inhibitor or vehiclecontrol for a final period of three days, after which myelination isvisualized by black gold staining (Millipore, Bedford, Mass.) followingthe manufacture's protocol. Images are acquired using a Leica M420microscope (Bannockburn, Ill.) and the staining intensity of corpuscallosum is analyzed using Metamorph software (Molecular Devices,Downingtown, Pa.). Three or four brain slices are used for eachtreatment group.

Lysolecithin Demyelination Model

Adult Sprague Dawley rats (220-260 g) are anesthetized byintraperitoneal injection of a cocktail, consisting of Ketamine (35mg/kg), Xylazine (6 mg/kg) and Acepromazine (1 mg/kg). The back of theanimal is shaved from the lower thoracic to the lumbar region,subsequently sanitized with 70% isopropanol, Betadine Scrub solution,and 70% isopropanol again. The animal is then placed onto stereotaxicframe.

After ensuring an adequate anesthetic level, the skin is incised alongthe midline over the thoracic region. The dorsal fascia is incised andthe paraspinal muscles separated from the spinous processes of thethoracic vertebrae T-9 through T-11. The T-10 vertebra is demolished,and the lamina removed with micro-rongeurs. Once the dorsal spinal cordregion is exposed, a microcapillary glass needle is inserted into thedorsal column to a depth of 0.6 mm. The demyelinating reagent, 1.5 μL of1% Lysolecithin (LPC, Sigma# L1381) in saline is injected with theinfusion rate of 2 nL/sec controlled by a micro-pump (World PrecisionInstrument #micro4). Once the injection is completed, the needle isplaced for additional 1 min before removal. The paraspinal muscles andthe lumbar fascia are closed with suture (#5, silk). The skin incisionis closed with wound clips. Animals are allowed to recover from theanesthesia and are observed in the humidified incubator.

Buprenorphine (0.05 mg/kg) is administrated subcutaneously (s.c.) twicea day for additional two days following operation.

Three days following the primary surgery, treatments with an S1P4receptor antagonist or ATX inhibitor (30 pmol), LPA (30 pmol) or control(0.1% DMSO in saline) are injected at the primary injection region in avolume of 1.5 μL with the same infusion speed as indicated above. Ninedays following the primary surgery, the animals are anesthetized andperfused trans-cardially with heparin (10 iu/mL) in saline followed by4% PFA in PBS. The spinal cords are removed and post fixed in PFAovernight. Then the cords are cut into 100 μM thickness longitudinallyand then 1% loxuol fast blue is stained and histological evaluation forremyelination and repair is assessed under microscope.

For systemic treatment, the animals are administered once dailyintraperitoneally with either an S1P4 receptor antagonist or ATXinhibitor (10 mg/kg) or control (15% HPCD(hydroxypropyl-β-cyclodextrin)) 2 days following the primary surgery.Nine days after the primary surgery, animals are sacrificed and thespinal cords were processed as indicated above.

In Vivo Screening Assays

Measurement of circulating lymphocytes: Compounds are dissolved in 30%HPCD. Mice (C57bl/6 male, 6-10 week-old) are administered 0.5 and 5mg/kg of a compound via oral gavage 30% HPCD is included as a negativecontrol.

Blood is collected from the retro-orbital sinus 5 and 24 hours afterdrug administration under short isoflurane anesthesia. Whole bloodsamples are subjected to hematology analysis. Peripheral lymphocytecounts are determined using an automated analyzer (HEMAVET™ 3700).Subpopulations of peripheral blood lymphocytes are stained byfluorochrome-conjugated specific antibodies and analyzed using afluorescent activating cell sorter (FACSCALIBUR™). Three mice are usedto assess the lymphocyte depletion activity of each compound screened.

Compounds of the invention can induce full lymphopenia at times as shortas 4 hours or less to as long as 48 hours or more; for example, 4 to 36hours, or 5 to 24 hours. In some cases, a compound of formula can inducefull lymphopenia at 5 hours and partial lymphopenia at 24 hours. Thedosage required to induce lymphopenia can be in the range of, e.g.,0.001 mg/kg to 100 mg/kg; or 0.01 mg/kg to 10 mg/kg. The dosage can be10 mg/kg or less, such as 5 mg/kg or less, 1 mg/kg or less, or 0.1 mg/kgor less.

CFA Inflammatory Pain Model

In the CFA (complete Freund's adjuvant) model, adult male SD (250-300 g)rats are anesthetized with isoflurane inhalation (4.5% induction/2.0%maintenance). Heat-killed M. Tuberculosis H37 RA (non-viable) suspendedat a concentration of 1.0 mg/ml in incomplete Freund's adjuvant is used(Chondrex Inc., catalog#7008). At day 0, intradermal injection (i.d.) of100 μl of CFA (1:1 oil/saline) is slowly perfused into the right footpadof the rats. At day 1, baseline tactile allodynia test are conducted:rats that develop sensitive painful response are enrolled to the study.At day 2, rats are orally dosed once with either vehicle or testcompound, then at 2 hrs, 4 hrs, 6 hrs and 24 hrs after dosage, all ratsare tested for mechanical allodynia response.

Tactile allodynia is tested as follows. A rat is placed in an elevatedPlexiglas observation chamber (approximately 4″×6″×10″) having a wiregrid (1 cm² spacing) mesh floor under polycarbonate cages. The rat isleft to acclimate to the experimental conditions for 20 minutes beforetesting begins. After the rat is calm, tactile allodynia is assessedusing a series of von Frey filaments ranging from 2.04-28.84 g(Stoelting, Wood Dale, Ill.). Graded pressure is presented to alocalized area on the plantar surface of the paw via the use of Von Freyhairs (monofilaments which are calibrated to bend at a known pressure).A response to the VonFrey hair is recorded as the rat withdrawing thetested paw and is usually followed by lifting and licking. A series offilaments are used to determine the threshold response using theestablished “Up-Down” method. Each paw is tested 4-6 times repeatedlywith 1-2 seconds (modified from Seltzer et al., 1991) in between eachprobe to accurately assess the behavior. A sharp lifting of the paw isscored as a positive response.

Rat Model of Neuropathic Pain

Chronic Constriction Injury (CCI) Surgery: In the CCI model (Bennett andXie, Pain, 1989, which is incorporated by reference in its entirety),adult male SD (250-275 g) rats are anesthetized with isofluraneinhalation (4.5% induction/2.0% maintenance). The surgery is performedunder aseptic conditions and involves exposing the sciatic nerve at themid-thigh level. Ocular lubricant is used as needed to prevent cornealdrying. After shaving and disinfecting the skin (betadine followed by70% ethanol), a small incision is made just caudal to the bicepsfemoris. Care is taken to not disturb the sciatic nerve. The nerve isslightly elevated, and 4 loose ligatures of 4-0 chromic gut suture areinserted under the nerve, and then are loosely tied around it. Thesutures constrict the nerve but do not strangle it. Prior to insertingthe chromic gut, it is rinsed twice in sterile saline. The incision isclosed with wound clips, and rats are allowed to recover from anesthesiaon a circulating water heating pad before being returned to their homecages. In the sham controls the skin is opened, and the sciatic nerve isidentified and elevated, but no sutures are tied around the nerve. Allrats are screened for pain response around post-surgery day 7 and onlyrats with sensitive pain response are enrolled to the study.

Animals are orally dosed twice/day for 3 times/week with either vehicleor test compound post-surgery at days 10, 12, 14, 17, 19 and 21, andanimals are also tested at the same schedule for three types ofneuropathic pain: thermal hyperalgesia, tactile allodynia andincapacitance.

(1) Plantar thermal hyperalgesia: Rats are tested for hyperalgesia usinga plantar device (Ugo Basile Inc., Cat.#37370). After acclimation to thetesting room, rats are placed on an elevated glass floor beneathinverted clear plastic cages, and a radiant heat source beneath theglass is aimed at the mid-plantar surface of the hindpaw after they haveceased all exploratory behavior. The onset of light activates a timer,which is terminated by a hindpaw withdrawal response. A cutoff time of30 seconds is used to avoid tissue damage in the absence of a response.The average withdrawal latency value of three trials from theipsilateral hindpaw is measured with at least 5-10 minutes between eachtrial to avoid any tissue damage.

(2) Tactile allodynia is tested as described above.

(3) Incapacitance: The incapacitance test measures the weight the ratplaces on each of its hindpaws. The rat is placed in a small, clearPlexiglas box (6″ long×3″ wide×4″ tall). The box is tilted up and opensin the front. The rat is placed in the box so that its hindpaws are atthe back (lower) portion of the box, and the forepaws are at the front(raised) part of the box. The rat's head is at the opening in the frontof the box. The box is placed on a divided scale such that each of therat's hindpaws is on one of the two weighing pans of the scale. Theweight that the rat placed on each hindpaw is then measured. Theprocedure is rapid (about 10 sec) and does not cause the animal anypain.

Other embodiments are within the scope of the following claims.

What is claimed is:
 1. A compound represented by formula (I):

or a pharmaceutically acceptable salt thereof, wherein: one of A⁴ and A⁵is CH and the other is CR⁴; R¹ is a C₆₋₂₀alkyl, or a cyclohexyl which isoptionally substituted with one R⁶; R⁴ is a group represented by thefollowing formula:

“

” represents the point of attachment; R⁶, for each occurrence, isindependently selected from the group consisting of C₁₋₆alkyl,C₁₋₆haloalkyl, C₃₋₈cycloalkyl and phenyl; R¹⁰ is hydrogen or aC₁₋₆alkyl; p is 0 or an integer from 1 to 6; and (i) m is 0; and R⁵ isselected from the group consisting of:

or (ii) m is 1; and R⁵ is cyclobutylene, cyclopentylene orcyclohexylene.
 2. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein the compound is represented bystructural formula (II):


3. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein the compound is represented by structural formula(III):


4. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein: m is 0; and R⁵ is selected from the group consistingof:


5. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein: m is 1; and R⁵ is cyclobutylene, cyclopentylene orcyclohexylene.
 6. A compound selected from the group consisting of:8-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[30.2.1]octane-3-carboxylicacid;9-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[30.3.1]nonane-3-carboxylicacid;3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)-2,2-dimethylcyclobutanecarboxylicacid;1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid;2-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)octahydrocyclopenta[c]pyrrole-5-carboxylicacid;1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)pyrrolidine-3-carboxylicacid;3-(((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)amino)cyclobutanecarboxylicacid;1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azepane-4-carboxylicacid;1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;1-((5-((cis-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;1-((5-((cis-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;1-((5-((cis-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;1-((5-((4,4-dimethylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;1-((5-((trans-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid;1-((5-((trans-4-phenylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid;1-((5-((trans-4-ethylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid;1-((5-((trans-4-(trifluoromethyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid;1-((5-((trans-4-isopropylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid;1-((5-((trans-4-butylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid;1-((5-((trans-4-cyclopentylcyclohexyl)oxy)naphthalen-2-yl)methyl)azetidine-3-carboxylicacid;1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid; 1-((5-(heptyloxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid;9-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-9-azabicyclo[3.3.1]nonane-3-carboxylicacid;8-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid;1-((5-((cis-4-methylcyclohexyl)oxy)naphthalen-2-yl)methyl)piperidine-4-carboxylicacid; 1-((5-(heptyloxy)naphthalen-1-yl)methyl)piperidine-4-carboxylicacid; and1-((5-((trans-4-(tert-butyl)cyclohexyl)oxy)naphthalen-1-yl)methyl)piperidine-4-carboxylicacid; or a pharmaceutically acceptable salt thereof.
 7. A pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier orexcipient and a compound according to claim 1, or a pharmaceuticallyacceptable salt thereof.
 8. A method of treating or reducing symptoms ofa condition selected from the group consisting of multiple sclerosis, achronic inflammatory disorder, asthma, an inflammatory neuropathy,arthritis, transplantation rejection, Crohn's disease, ulcerativecolitis, lupus erythematosis, psoriasis, an ischemia-reperfusion injury,a solid tumor, a tumor metastasis, a vascular disease, a pain condition,an acute viral disease, an inflammatory bowel condition,insulin-dependent diabetes, non-insulin dependent diabetes, a fibrosisof the lung, and a malignancy of the lung in a mammal comprisingadministering to said mammal an effective amount of a compound accordingto claim 1, or a pharmaceutically acceptable salt thereof.
 9. The methodof claim 8, wherein the condition is multiple sclerosis.
 10. The methodof claim 8, wherein the condition is rheumatoid arthritis.
 11. Themethod of claim 8, further comprising administering to said mammal aneffective amount of one or more drugs selected from the group consistingof: a corticosteroid, a bronchodilator, an antiasthmatic, anantiinflammatory, an antirheumatic, an immunosuppressant, anantimetabolite, an immunomodulating agent, an antipsoriatic, and anantidiabetic.
 12. A method of treating or reducing chronic pain in amammal comprising administering to said mammal an effective amount of acompound according to claim 1, or a pharmaceutically acceptable saltthereof.
 13. The method of claim 12, wherein the chronic pain isinflammatory pain.
 14. The method of claim 13, wherein the chronic painis neuropathic pain.